Mar. 2016, 日本蚕糸学会, 日本蚕糸学会賞　　　　　　　　　　　　　　　
浅野 眞一郎

Whole Genome Resequencing Reveals Origins and Global Invasion Pathways of the Japanese Beetle Popillia japonica.
Rebecca Funari, Elahe Parvizi, Claudio Cucini, Sara Boschi, Elena Cardaioli, Daniel A Potter, Shin-Ichiro Asano, Duarte Toubarro, Luca Jelmini, Francesco Paoli, Antonio Carapelli, Angela McGaughran, Francesco Frati, Francesco Nardi
Molecular ecology, 34, 16, e70008, Aug. 2025, [Peer-reviewed], [International Magazine]
English, Scientific journal, Invasive species are an increasing global threat given their ability to rapidly spread and adapt to novel environments. The adverse ecological and economic impacts of invasive species highlight the critical need to understand the mechanisms that underpin invasion processes and success. The Japanese beetle, Popillia japonica, is an invasive pest of remarkable interest, as it feeds on hundreds of economically valuable plant species. It has been expanding outside of its native range in Japan since the first decades of the 20th century, colonising large areas of North America and, more recently, Europe. Here, we compared whole-genome resequencing data from individuals encompassing the entire species distribution to study the geographic differentiation of P. japonica populations and reconstruct expansion routes from Japan to the USA and Europe. We found six genomically distinguishable clusters, corresponding to the approximate colonisation areas at a continental scale. Our analysis supported an ancestral divergence between South and North/Central Japan, with the latter being the source of the initial invasion to the USA. Coalescent simulations supported independent bridgehead events from the USA to the Azores and Italy. We also investigated possible signals of selection to better understand the adaptive mechanisms that underlie the invasion success of P. japonica. However, the absence of strong selection signatures suggested that the beetle's adaptive ability might be embedded in pre-existing genomic features. Our comprehensive genome-wide dataset allowed a detailed inference of the invasion process and may be useful in determining the origin of P. japonica individuals in future invasion events.

Functional expression of Bacillus thuringiensis Cry proteins on the surface of Bacillus subtilis spores.
Sachiko Kashojiya, Shin-Ichiro Asano, Paul W Oeller, Takashi Yamamoto
Journal of pesticide science, 49, 4, 232, 242, 20 Nov. 2024, [Peer-reviewed], [Domestic magazines]
English, Scientific journal, The Cry1Fa insecticidal protein from Bacillus thuringiensis (Bt) was expressed on the surface of Bacillus subtilis (Bs) spores to create transgenic Bs spores referred to as Spore-Cry1Fa. Cry1Fa, along with its leader sequence, was connected to the carboxyl end of a Bs spore outercoat protein, CotC, through a flexible linker. The Arg-27 residue of the Cry1Fa protein was mutated to Leu to prevent detachment from the spores due to protease digestion. The expression of the Cry protein on the Bs spores was confirmed by fluorescence microscopy using an anti-Cry1Fa antibody. The Cry protein, tightly anchored to the spore surface, appeared to be functional in terms of receptor binding. Spore-Cry1Fa bound to Sf9 cells expressing Spodoptera frugiperda (Sf) ABCC2 transporter and killed the cells within 60 min. Additionally, nano-lipid particles of SfABCC2 were produced using styrene-maleic acid (SMA) to demonstrate the binding to Spore-Cry1Fa.

Clinically approved chemical-controlled suppression of protein expression in BmN cells
Toshiki Nakanishi, Shin-ichiro Asano, Hisanori Bando, Masanao Sato
Journal of Insect Biotechnology and Sericology, 93, 1, 1, 10, 28 May 2024, [Peer-reviewed]
English, Scientific journal

The direction, timing and demography of Popillia japonica (Coleoptera) invasion reconstructed using complete mitochondrial genomes.
Francesco Nardi, Sara Boschi, Rebecca Funari, Claudio Cucini, Elena Cardaioli, Daniel Potter, Shin-Ichiro Asano, Duarte Toubarro, Michela Meier, Francesco Paoli, Antonio Carapelli, Francesco Frati
Scientific reports, 14, 1, 7120, 7120, 26 Mar. 2024, [International Magazine]
English, Scientific journal, The Japanese beetle Popillia japonica is a pest insect that feeds on hundreds of species of wild and cultivated plants including important fruit, vegetable, and field crops. Native to Japan, the pest has invaded large areas of the USA, Canada, the Azores (Portugal), Italy, and Ticino (Switzerland), and it is considered a priority for control in the European Union. We determined the complete mitochondrial genome sequence in 86 individuals covering the entire distribution of the species. Phylogenetic analysis supports a major division between South Japan and Central/North Japan, with invasive samples coming from the latter. The origin of invasive USA samples is incompatible, in terms of the timing of the event, with a single introduction, with multiple Japanese lineages having been introduced and one accounting for most of the population expansion locally. The origin of the two invasive European populations is compatible with two different invasions followed by minimal differentiation locally. Population analyses provide the possibility to estimate the rate of sequence change from the data and to date major invasion events. Demographic analysis identifies a population expansion followed by a period of contraction prior to the invasion. The present study adds a time and demographic dimension to available reconstructions.

Proliferation of Bombyx mori nucleopolyhedrovirus strain H4 in BmN cells is enhanced by exchange of the F gene sequence with type strain T3
Mami Sakai, Satoshi Kakutani, Shin-ichiro Asano, Masanao Sato, Hisanori Bando
Virus Research, 291, 198195, 198195, Elsevier BV, Jan. 2021, [Peer-reviewed], [International Magazine]
English, Scientific journal, The Bombyx mori nucleopolyhedrovirus (BmNPV)-based baculoviral expression vector system is among the most efficient expression vector systems for eukaryotic proteins especially when used in combination with silkworms as a host. We newly isolated a novel BmNPV strain (BmNPV H4) in Hokkaido, Japan that outperforms the type strain T3 in terms of both proliferation and expression of polyhedrin protein in silkworm larvae; however, it proliferates poorly in the BmN cell line. We inferred the gene responsible for the differences in proliferation between viral strains by quantifying amino acid similarity distances in protein functional domains and identifying highly divergent alleles between the H4 and T3 strains. Among proteins that differ markedly in functional domain sequence between H4 and T3, we identified the F gene, which encodes the F protein, as a putative cause of proliferative differences between the two strains. Using recombinant viruses with the F protein-coding sequence exchanged between H4 and T3, we determined that the T3 F protein increases H4 proliferation in BmN while the H4 F protein does not improve T3 proliferation in silkworm larvae. Our results suggest that the BmNPV F protein can strongly affect viral proliferation in a genetic background-specific manner and may be an important target for manipulating the proliferation characteristics of BmNPV-based expression vectors.

A single amino acid replacement of tyrosine with histidine at position 172 in BmNPV T3 GP64 decreases viral proliferation in silkworm larvae despite increasing membrane fusion activity with BmN cells　　　　　　　　　　　　　　　
Mari Sekiguchi, Sayuri Ishikawa, Shin-ichiro Asano, Hisanori Bando, Masanao Sato
Journal of Insect Biotechnology and Sericology, 89, 3, 45, 53, Sep. 2020, [Peer-reviewed]
Scientific journal

Detection of cry-type gene homologues in the draft genome sequence of paenibacillus popilliae ATCC 14706t and trial for the expression of recombinant proteins in escherichia coli
Akio Kawahara, Kazuhiro Iiyama, Shin-Ichiro Asano, Oumi Nishi, Chisa Yasunaga-Aoki
Journal of Insect Biotechnology and Sericology, 89, 2, 25, 30, Japanese Society of Sericultural Sciences, 2020, [Peer-reviewed]
English, Scientific journal

Design and prototyping of a Bombyx mori nucleopolyhedrovirus-based genome assembly from PCR-amplified DNA fragments　　　　　　　　　　　　　　　
Naoto Harada, Daiki Ishibashi, Shin-ichiro Asano, Masanao Sato, Hisanori Bando
Insect Biotechnology and Sericology, 87, 3, 97, 108, Oct. 2018, [Peer-reviewed]
English, Scientific journal

Bombyx mori nucleopolyhedrovirus immediate‐early遺伝子ORF上流500塩基による発現制御についての解析
工藤滉己, 佐藤昌直, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 42, 8‐11, 25 Dec. 2017
Japanese

BmNPVゲノムのin vitroアセンブリ
原田直人, 佐藤昌直, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 42, 12‐16, 25 Dec. 2017
Japanese

マメコガネ幼虫におけるCry8Daトキシンの殺虫活性機構について
山口真, 佐藤昌直, 伴戸久徳, 浅野眞一郎
東北蚕糸・昆虫利用研究報告, 42, 1‐3, 25 Dec. 2017
Japanese

バキュロウイルスの複数の非必須遺伝子がウイルス増殖に与える影響について
齋藤諒, 武田ひとみ, 佐藤昌直, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 42, 4‐7, 25 Dec. 2017
Japanese

Cry8Daトキシンの殺虫活性機構について
池田優里恵, 佐藤昌直, 伴戸久徳, 浅野眞一郎
東北蚕糸・昆虫利用研究報告, 41, 12‐13, 10 Dec. 2016
Japanese

Cry1Iaトキシンのコナガに対する殺虫活性機構について
佐藤文美, 佐藤昌直, 伴戸久徳, 浅野眞一郎
東北蚕糸・昆虫利用研究報告, 41, 7‐8, 10 Dec. 2016
Japanese

Generation of an infectious cDNA clone of Okushiri virus and its derivative capable of expressing an exogenous gene
KURNIA Yudistira Wahyu, FUJITA Ryosuke, SATO Masanao, ISAWA Haruhiko, ASANO Shin‐ichiro, BINH Ngo Dinh, BANDO Hisanori
Journal of Insect Biotechnology and Sericology, 85, 2, 39‐47, 12 Sep. 2016
English

Characterization of a novel negevirus isolated from Aedes larvae collected in a subarctic region of Japan
Kota Kawakami, Yudistira Wahyu Kurnia, Ryosuke Fujita, Toshiaki Ito, Haruhiko Isawa, Shin-ichiro Asano, Ngo Dinh Binh, Hisanori Bando
ARCHIVES OF VIROLOGY, 161, 4, 801, 809, Apr. 2016, [Peer-reviewed]
English, Scientific journal

BmSyndecanノックダウンBmN細胞におけるBmNPVの増殖
宇多桃香, 佐藤昌直, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 40, 12, 14, 20 Dec. 2015
Japanese

バキュロウイルスゲノム人工合成系の確立
石橋大樹, 佐藤昌直, 高ひとみ, 武内潤一, 浅野眞一郎, 伴戸久徳
日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 85th, 58, 25 Sep. 2015
Japanese

Analysis of the Bombyx mori nucleopolyhedrovirus ie-1 promoter in insect, mammalian, plant, and bacterial cells
Ryosuke Fujita, Chikako Ono, Isamu Ono, Shin-ichiro Asano, Hisanori Bando
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 464, 4, 1297, 1301, Sep. 2015, [Peer-reviewed]
English, Scientific journal

Characterization of KfrA proteins encoded by a plasmid of Paenibacillus popilliae ATCC 14706^T
Kazuhiro Iiyama, Hiroaki Mon, Kazuki Mori, Takumi Mitsudome, Jae Man Lee, Takahiro Kusakabe, Kousuke Tashiro, Shin-ichiro Asano, Chisa Yasunaga-Aoki
Meta Gene, 4, 29, 44, Elsevier, 01 Jun. 2015, [Peer-reviewed]
English, Scientific journal

Tightly Regulated Expression of Autographa californica Multicapsid Nucleopolyhedrovirus Immediate Early Genes Emerges from Their Interactions and Possible Collective Behaviors
Chikako Ono, Masanao Sato, Hitomi Taka, Shin-ichiro Asano, Yoshiharu Matsuura, Hisanori Bando
PLOS ONE, 10, 3, e0119580, Mar. 2015, [Peer-reviewed]
English, Scientific journal

バキュロウイルスゲノム人工合成系の確立に向けて
石橋大樹, 佐藤昌直, 高ひとみ, 武内潤一, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 39, 5, 7, 10 Dec. 2014
Japanese

Phylogenetic relationship of Paenibacillus species based on putative replication origin regions and analysis of an yheCD-like sequence found in this region
Kazuhiro Iiyama, Masahiro Otao, Kazuki Mori, Hiroaki Mon, Jae Man Lee, Takahiro Kusakabe, Kousuke Tashiro, Shin-Ichiro Asano, Chisa Yasunaga-Aoki
BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, 78, 5, 891, 897, May 2014, [Peer-reviewed]
English, Scientific journal

Transcriptional profile of tomato roots exhibiting Bacillus thuringiensis-induced resistance to Ralstonia solanacearum
Hideki Takahashi, Kazuhiro Nakaho, Takeaki Ishihara, Sugihiro Ando, Takumi Wada, Yoshinori Kanayama, Shinichiro Asano, Shigenobu Yoshida, Seiya Tsushima, Mitsuro Hyakumachi
PLANT CELL REPORTS, 33, 1, 99, 110, Jan. 2014, [Peer-reviewed], [International Magazine]
English, Scientific journal

BmNPVの非必須遺伝子(Bm32‐Bm36)間相互作用解析
高ひとみ, 小野慎子, 佐藤昌直, 浅野眞一郎, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 38, 1, 3, 30 Dec. 2013
Japanese

Mos1 transposon-based transformation of fish cell lines using baculoviral vectors
Masako Yokoo, Ryosuke Fujita, Yumiko Nakajima, Mamoru Yoshimizu, Hisae Kasai, Shin-ichiro Asano, Hisanori Bando
Biochemical and Biophysical Research Communications, 439, 1, 18, 22, 13 Sep. 2013, [Peer-reviewed]
English, Scientific journal

Putative loop 1 residues in domain II of Cry39Aa toxin are important for larvicidal activity against Anopheles stephensi
ISHIGAKI Shun-ichiro, BANDO Hisanori, ASANO Shin-ichiro
The Journal of Sericultural Science of Japan, 82, 1, 13, 18, The Japanese Society of Sericultural Science, 28 Jun. 2013
English, Cry39Aa from Bacillus thuringiensis serovar aizawai BUN1-14 is highly toxic to the larvae of Anopheles stephensi mosquitoes, which transmit malarial parasites. We constructed a homology model of Cry39Aa toxin using the reported structure of Cry4Ba toxin (PDB file 1W99) as a template, and we identified 349KYAYWR354 as the putative loop 1 in domain II. Many studies of various Cry toxins have shown that surface-exposed loops of domain II are critical for toxicity and receptor binding. To investigate the functional role of the putative loop 1 of Cry39Aa toxin, we performed site-directed mutagenesis in the loop. The results of alanine substitutions revealed that the entire structure of loop 1, and aromatic amino acids Y350 and Y352 in particular, are essential for larvicidal activity. In contrast, the toxicities of Cry39Aa mutants with phenylalanine substitutions at these two tyrosine residues did not differ markedly from the toxicity of wild-type Cry39Aa. A competitive binding assay involving A. stephensi brush border membrane vesicles revealed that the alanine mutants showed reduced competition with wild-type Cry39Aa toxin. Our results suggest that the molecular structure of loop 1 in domain II of Cry39Aa toxin is important for toxicity and receptor binding in the midgut of A. stephensi larvae.

Complex genetic interactions among non-essential genes of BmNPV revealed by multiple gene knockout analysis
TAKA Hitomi, ONO Chikako, SATO Masanao, ASANO Shin‐ichiro, BANDO Hisanori
J Insect Biotechnol Sericology, 82, 1, 25, 32, 28 Jun. 2013
English, Scientific journal

Identification of a Bacillus thuringiensis Cry8Da toxin-binding glucosidase from the adult Japanese beetle, Popillia japonica
Takuya Yamaguchi, Hisanori Bando, Shin-ichiro Asano
JOURNAL OF INVERTEBRATE PATHOLOGY, 113, 2, 123, 128, Jun. 2013, [Peer-reviewed], [International Magazine]
English, Scientific journal

Bacillus thuringiensis suppresses bacterial wilt disease caused by Ralstonia solanacearum with systemic induction of defense-related gene expression in tomato
Mitsuro Hyakumachi, Mitsuyoshi Nishimura, Tatsuyuki Arakawa, Shinichiro Asano, Shigenobu Yoshida, Seiya Tsushima, Hideki Takahashi
Microbes and Environments, 28, 1, 128, 134, 2013, [Peer-reviewed], [Domestic magazines]
English, Scientific journal

Phylogenetic analysis of Paenibacillus popilliae and its related taxa based on housekeeping genes
Kazuhiro Iiyama, Oumi Nishi, Hiroaki Mon, Jae Man Lee, Takahiro Kusakabe, Shin-ichiro Asano, Chisa Yasunaga-Aoki, Susumu Shimizu
Journal of Insect Biotechnology and Sericology, 82, 1, 1, 11, The Japanese Society of Sericultural Science, 2013, [Peer-reviewed]
English, Scientific journal, Four housekeeping genes including gapA, groEL, gyrA and pgi were used in order to reveal a phylogenetic relationship among Paenibacillus species. Phylogenetic analysis and end-to-end pairwise analysis were carried out using nucleotide and amino acid sequences of the housekeeping genes. A monophylogenetic clade including P. popilliae, P. thiaminolyticus and P. dendritiformis formed in all phylogenetic trees. The results in both analyses indicated that the relationship between P. larvae subsp. larvae and P. popilliae was comparatively far. The result of the end-to-end pairwise analysis also showed that, among the Paenibacillus species used in this study, the closest species to P. popilliae was P. dendritiformis.

Draft Genorne Sequence of Paenibacillus popilliae ATCC 14706(T)
liyama Kazuhiro, Mori Kazuki, Mon Hiroaki, Chieda Yuuka, Lee Jae Man, Kusakabe Takahiro, Tashiro Kousuke, Asano Shin-Ichiro, Yasunaga-Aoki Chisa, Shimizu Susumu
Journal of Insect Biotechnology and Sericology, 82, 2, 45, 48, 2013, [Peer-reviewed]

A 50-Kilodalton Cry2A Peptide Is Lethal to Bombyx mori and Lymantria dispar
Masataka Ohsawa, Miki Tanaka, Kenta Moriyama, Mitsuaki Shimazu, Shin-ichiro Asano, Kazuhisa Miyamoto, Kohsuke Haginoya, Toshiaki Mitsui, Tomoaki Kouya, Masayuki Taniguchi, Hidetaka Hori
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 78, 13, 4755, 4757, Jul. 2012, [Peer-reviewed]
English, Scientific journal

An HDAC inhibitor increases AcMNPV gene expression in mammalian cells
Ryosuke Fujita, Daisuke Ohtsuka, Ken Sahara, Shinichiro Asano, Hisanori Bando
ARCHIVES OF VIROLOGY, 155, 4, 577, 581, Apr. 2010, [Peer-reviewed]
English, Scientific journal

Supression of AcMNPV gene expression in mammalian cells　　　　　　　　　　　　　　　
Fujita R, Asano S, Sahara K, Bando H
41th Annual meeting of the society for invertebrate pathology and 9th international conference on Bacillus thuringiensis, 19, 19, Aug. 2008, [Peer-reviewed]
English, International conference proceedings

Ecdysone response element in a baculovirus immediate-early gene, ie1, promoter
K. Kojima, K. Kojima, K. Oritani, T. Nakatsukasa, S. Asano, K. Sahara, H. Bando
Virus Research, 130, 1, 202, 209, Elsevier, 01 Dec. 2007
English

Restricted gene transcription of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) in mammalian cells　　　　　　　　　　　　　　　
Fujita R, Asano S, Sahara K, Bando H
40th Annual meeting of the society for invertebrate pathology and 1st international forum on entomopathogenic nematodes and symbiotic bacteria, 95, 95, Aug. 2007, [Peer-reviewed]
English, International conference proceedings

Expression of Autographa californica multiple nucleopolyhedrovirus genes in mammalian cells and Upregulation of the host beta-actin gene
R Fujita, T Matsuyama, J Yamagishi, K Sahara, S Asano, H Bando
JOURNAL OF VIROLOGY, 80, 5, 2390, 2395, Mar. 2006, [Peer-reviewed]
English, Scientific journal

Enhancement of cauliflower mosaic virus 35S promoter in insect cells infected with baculovirus
Abe T, Miyake N, Nishijima Y, Fujita R, Sahara K, Asano S, Bando H
Virus Research, 112, 1-2, 38, 41, Oct. 2005, [Peer-reviewed]
English, Scientific journal

昆虫病原ウイルスAcNPVの哺乳動物細胞における遺伝子発現とその影響　　　　　　　　　　　　　　　
藤田龍介, 浅野眞一郎, 佐原健, 伴戸久徳
東北蚕糸・昆虫利用研究報告, 29, 10, 10, Dec. 2004, [Peer-reviewed]
Japanese, Research society

Resistance of transgenic silkworm against Bombyx mori nucleopolyhedrovirus (BmNPV)
小島 桂, 斎藤 寛, 山田 恭裕, 田村 俊樹, 磯部 良子, 浅野 眞一郎, 佐原 健, 伴戸 久徳
Research bulletin of the University Farm, Hokkaido University, 33, 9-13, 33, 9, 13, 北海道大学北方生物圏フィールド科学センター耕地圏ステーション生物生産研究農場, Mar. 2003
Japanese, Transgenic silkworms possessing artificial anti-viral genes were constructed using a transposon, piggyBac, system and subjected to test for their resistance to Bombyx mori nucleopolyhedrovirus (BmNPV). Silkworms inoculated with BmNPV at the 1st day of the 2nd instar larvae survived till 4th instar larvae, and all of them were found to be transgenic silkworms. Though apparent BmNPV resistance increased in the transgenic silkworms, further improvement of the resistance was required for practical use.

Use of an N-terminal half truncated IE1 as an antagonist of IE1, an essential regulatory protein in baculovirus
Y Yamada, T Matsuyama, GX Quan, T Kanda, T Tamura, K Sahara, S Asano, H Bando
VIRUS RESEARCH, 90, 1-2, 253, 261, Dec. 2002
English, Scientific journal

Antisense and double-strand RNA interference in a silkworm ovarian cell line　　　　　　　　　　　　　　　
Ryoko Isobe, Katsura Kojima, Ken Sahara, Shin-ichiro Asano, Hisanori Bando
Journal of Insect Biotechnology and Sericology, 71, 1, 43, 47, 2002
English, Scientific journal

Comparative results of wild silkworm cocoon, Antheraea yamamai, reared with artificial diet during younger larval stage.
斎藤 寛, 菊池 邦夫, 山田 恭裕, 飯塚 敏彦, 浅野 眞一郎, 伴戸 久徳, 佐原 健
Research bulletin of the University Farm,Hokkaido University, 32, 61-69., 32, 61, 69, 北海道大学農学部附属農場, Mar. 2001
Japanese

Analysis of proteins encoded in the bipartite genome of a new type of parvo-like virus isolated from silkworm - structural protein with DNA polymerase motif
T Hayakawa, K Kojima, K Nonaka, M Nakagaki, K Sahara, S Asano, T Iizuka, H Bando
VIRUS RESEARCH, 66, 1, 101, 108, Jan. 2000, [Peer-reviewed]
English, Scientific journal

Analysis of genetic information of an insect picorna-like virus, infectious flacherie virus of silkworm: Evidence for evolutionary relationships among insect, mammalian and plant picorna(-like) viruses
H. Isawa, S. Asano, K. Sahara, T. Iizuka, H. Bando, H. Bando
Archives of Virology, 143, 127-143, 127, 143, 27 Mar. 1998

Characterization of a cry2A gene cloned from an isolate of Bacillus thuringiensis serovar sotto
J Sasaki, S Asano, N Hashimoto, BW Lay, S Hastowo, H Bando, T Iizuka
CURRENT MICROBIOLOGY, 35, 1, 1, 8, Jul. 1997, [Peer-reviewed]
English, Scientific journal

Cloning of novel enterotoxin genes from Bacillus cereus and Bacillus thuringiensis
S Asano, Y Nukumizu, H Bando, T Iizuka, T Yamamoto
APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 63, 3, 1054, 1057, Mar. 1997, [Peer-reviewed]
English, Scientific journal

Detection of replicative intermediate with closed terminus of Bombyx densonucleosis virus - Brief report
T Hayakawa, S Asano, K Sahara, T Iizuka, H Bando
ARCHIVES OF VIROLOGY, 142, 2, 393, 399, 1997, [Peer-reviewed]
English, Scientific journal

Insecticidal activity of the protein encoded by the cryV gene of Bacillus thuringiensis kurstaki INA-02
J Sasaki, S Asano, T Iizuka, H Bando, BW Lay, S Hastowo, GK Powell, T Yamamoto
CURRENT MICROBIOLOGY, 32, 4, 195, 200, Apr. 1996, [Peer-reviewed]
English, Scientific journal

Analysis of the genetic information of a DNA segment of a new virus from silkworm
H. Bando, T. Hayakawa, S. Asano, K. Sahara, M. Nakagaki, T. Iizuka
Archives of Virology, 140, 1147-1155, 6, 1147, 1155, Springer Science and Business Media LLC, 01 Jun. 1995
Scientific journal

北海道の土壌から分離したBacillus thuringiensis株の性状
佐々木 潤, 浅野 真一郎, 伴戸 久徳
日本蚕糸学雑誌, 63, 5, p361, 366, 日本蠶絲學會, Oct. 1994, [Peer-reviewed]
Japanese

CryIA(b) gene from Bacillus thuringiensis subsp. wuhanensis; DNA sequence analysis and insecticidal activity
Ikuhiro Nakatani, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka
Journal of Sericultural Science of Japan, 63, 2, 140, 148, 1994, [Peer-reviewed]
English, Scientific journal

Insecticidal activity of the cryIA(a) and cryIB delta-endotoxin of Bacillus thuringiensis was retained after the coding region of the gene was truncated and expressed in Escherichia coli
Toshihiko Iizuka, Seiichiro Ishikawa, Shinichiro Asano, Hisanori Bando, Zhenwei Zheng, Norimoto Murai
Journal of Sericultural Science of Japan, 63, 4, 303, 309, 1994, [Peer-reviewed]
English, Scientific journal

Transplanting the Human TBX5 Regulatory Elements: In Silico Screening and Evaluation of Transcription Factors Independent of Bombyx mori Transcriptional Regulation
前田麻亜子, 坪田拓也, 門宏明, 増田亮津, 松田拓也, 浅野眞一郎, 伴戸久徳, 瀬筒秀樹, 瀬筒秀樹, 荒川和晴, 荒川和晴, 日下部宜宏, 佐藤昌直, 佐藤昌直, 日本分子生物学会年会プログラム・要旨集(Web), 46th, 2023

蛍光レポーター発現細胞クラスター認識を自動化・定量化する画像解析アプローチ
中西登志紀, 関口真理, 浅野眞一郎, 伴戸久徳, 佐藤昌直, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 90th, 2020

グアム産タイワンカブトにおけるOrNV配列の調査
橋本健太郎, 佐藤昌直, 伴戸久徳, 浅野眞一郎, 東北蚕糸・昆虫利用研究報告, 45, 2020

BmNPVにおける膜タンパク質(GP64)のアミノ酸バリアントが増殖に与える影響
関口真理, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 90th, 2020

カイコ核多角体病ウイルス新規株H4の膜タンパク質GP64の機能解析
関口真理, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 89th, 2019

BmN細胞におけるSmall Molecule Assisted Shutoffの検証
中西登志紀, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 89th, 2019

ネッタイシマカにおけるCry44Aaトキシンレセプターの評価
浅野眞一郎, 小野山雄亮, 中神あゆみ, 中尾悠太, 佐藤昌直, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 88th, 2018

BmNPV非必須遺伝子arif-1,pif-2による補償的な必須遺伝子発現制御
高ひとみ, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 87th, 2017

ネッタイシマカにおけるCry44Aaトキシンレセプターについて
浅野眞一郎, 中神あゆみ, 中尾悠太, 佐藤昌直, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 87th, 2017

Ecology of Entomopathogenic Bacillus thuringiensis(1)Bacillus thuringiensis in Phytobiome
小池 正徳, 高橋 尚樹, 笹原 勇太, 斉佳 鶴怜, 石井 嶺広, 相内 大吾, 浅野 眞一郎, 昆虫と自然, 51, 13, 39, 41, Dec. 2016
ニューサイエンス社, Japanese

Dual-control using insect pathogen
小池 正徳, 斉 佳鶴玲, 相内 大吾, 石井 嶺広, 浅野 眞一郎, 昆虫と自然, 51, 7, 45, 48, Jun. 2016
ニューサイエンス社, Japanese

Insect pathogenic bacteria
浅野 眞一郎, 昆虫と自然, 51, 3, 4, 7,図巻頭1p, Mar. 2016
ニューサイエンス社, Japanese

BmNPVのアクセサリー遺伝子が担うウイルス増殖のロバストネス
高ひとみ, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本分子生物学会年会プログラム・要旨集(Web), 39th, 2016

必須性を示すBmNPV非必須遺伝子クラスターの解析
高ひとみ, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 85th, 2015

BmSyndecanノックダウンBmN細胞におけるBmNPVの増殖
宇多桃香, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 85th, 2015

AcMNPV IE遺伝子発現のロバストネスを支えるIE遺伝子ネットワークと補償的相互作用
小野慎子, 高ひとみ, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 84th, 70, 09 Mar. 2014
Japanese

Cry2Aの殺虫活性について
高野恵美子, 高橋こう, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 84th, 70, 09 Mar. 2014
Japanese

非必須遺伝子の複数ノックアウトがBmNPV増殖に与える影響について
高ひとみ, 小野慎子, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 84th, 2014

非必須遺伝子の多重ノックアウトがBmNPV増殖に与える影響
高ひとみ, 小野慎子, 佐藤昌直, 浅野眞一郎, 伴戸久徳, 日本分子生物学会年会プログラム・要旨集(Web), 37th, 2014

コクヌストモドキ由来A Disintegrin And Metalloprotease(ADAM)について
半谷大輝, 伴戸久徳, 浅野眞一郎, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 83rd, 64, 18 Mar. 2013
Japanese

ハマダラカ由来アルカリフォスファターゼのCry39Aaトキシン受容体としての機能解析
石垣俊一郎, 伴戸久徳, 浅野眞一郎, 日本応用動物昆虫学会大会講演要旨, 57th, 190, 15 Mar. 2013
Japanese

昆虫特異的フラビウイルスAedes flavivirus(AEFV)ベトナム分離株の遺伝子構造解析ならびにAEFVの伝播様式の解析
小林大介, 伊澤晴彦, 鍬田龍星, 星野啓太, BINH Ngo Dinh, 浅野眞一郎, 伴戸久徳, 糸山享, 沢辺京子, 日本応用動物昆虫学会大会講演要旨, 57th, 5, 15 Mar. 2013
Japanese

Cry2Aaの蚊類殺虫活性におけるDomain II loop領域の機能解析
高橋瑛, 伴戸久徳, 浅野眞一郎, 日本応用動物昆虫学会大会講演要旨, 57th, 190, 15 Mar. 2013
Japanese

Phenotypic grouping of 141 BmNPVs lacking viral gene sequences
Chikako Ono, Takanori Kamagata, Hitomi Taka, Ken Sahara, Shin-ichiro Asano, Hisanori Bando, VIRUS RESEARCH, 165, 2, 197, 206, May 2012
English

Aedes類蚊幼虫より分離した新規植物様ウイルスの解析
川上広太, WAHYU Kurnia Yudistira, 伊澤晴彦, BINH Ngo Dinh, 浅野眞一郎, 伴戸久徳, 東北蚕糸・昆虫利用研究報告, 37, 2012

B.thuringiensis92‐10株の産生するParasporin‐1Aaについて
上森翔太, 浅野眞一郎, 佐原健, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 81st, 72, 20 Mar. 2011
Japanese

蚊由来培養細胞に対し毒性を示すウイルスの単離及び同定
川上広太, 伊澤晴彦, 伊藤利章, 浅野眞一郎, BINH Ngo Dinh, 佐原健, 伴戸久徳, 東北蚕糸・昆虫利用研究報告, 35/36, 2011

Isolation of the Thosea asigna virus (TaV) from the epizootic Setothosea asigna larvae collected in South Sumatra and a study on its pathogenicity to Limacodidae larvae in Japan
Maria Sugiharti, Chikako Ono, Shin-Ichiro Asano, Ken Sahara, Hisanori Bando, Toshiaki Ito, Yulia Pujiastuti, Journal of Biotechnology and Sericology, 79, 3, 117, 124, 2011
English

Possibility of insect pathogenic microbe as hygiene insect pest control materials
Shin-ichiro Asano, JOURNAL OF PESTICIDE SCIENCE, 36, 1, 91, 93, 2011
Japanese, Book review

衛生害虫防除資材としての昆虫病原微生物の可能性
浅野眞一郎, 日本農薬学会誌, 36, 1, 91, 93, 2011

Intramolecular proteolytic nicking and binding of Bacillus thuringiensis Cry8Da toxin in BBMVs of Japanese beetle
Takuya Yamaguchi, Ken Sahara, Hisanori Bando, Shin-ichiro Asano, JOURNAL OF INVERTEBRATE PATHOLOGY, 105, 3, 243, 247, Nov. 2010
English

2S-2 Possibility of insect pathogenic microbe as hygiene insect pest control materials
ASANO Shin-ichiro, 講演要旨集, 35, 34, 34, 21 Apr. 2010
Pesticide Science Society of Japan, Japanese

蚕糸昆虫研究におけるＢＴ研究の歴史について
浅野 眞一郎, 蚕糸・昆虫バイオテック, 77, 3, 181, 186, 2009
社団法人 日本蚕糸学会, Japanese

蚕糸昆虫研究におけるBT研究の歴史について
浅野 眞一郎, 蚕糸・昆虫バイオテック = Sanshi-konchu biotec, 77, 3, 181, 186, 01 Dec. 2008
Bacillus thuringiensis（Bt）は1901年にわが国の石渡繁胤によって、カイコに卒倒病を引き起こす病原細菌として、世界に先駆けて報告された。しかしながら、論文が日本語で書かれていたこと、新種の微生物として記載を行わなかったことから、注目を集めなかった。1911年にはBerlinerがドイツのThuringiaの地で、ノシメマダラメイガの斃死幼虫から分離した昆虫病原細菌を、発見地にちなみBacillus thuringiensisをいう学名を与えた。日本国内とりわけ蚕糸昆虫研究者の間では、B. thuringiensisがわが国の石渡によって発見されたsotto菌であることは知られていたが、世界においてこの菌の発見者が石渡であると認識されるようになったのは、その後約半世紀近く経ってからだった。Bt研究については、石渡が本菌の発見者であることからも分かるように、これまでの最先端の研究がわが国の蚕糸昆虫研究者によってなされてきた。ここでは、蚕糸昆虫研究におけるB. thuringiensis研究の歴史と今後の研究について、本特集でとりあげた最近の研究を含め紹介したい。, 日本蚕糸学会, Japanese

Discovery of a novel Bacillus thuringiensis Cry8D protein and the unique toxicity of the Cry8D-class proteins against scarab beetles
Takuya Yamaguchi, Ken Sahara, Hisanori Bando, Shin-ichiro Asano, JOURNAL OF INVERTEBRATE PATHOLOGY, 99, 3, 257, 262, Nov. 2008
English

Use of Bombyx mori U6 Promoter for Inducing Gene-Silencing in Silkworm Cells
OHTSUKA Daisuke, NAKATSUKASA Tomonori, FUJITA Ryosuke, ASANO Shin-ichiro, SAHARA Ken, BANDO Hisanori, Journal of insect biotechnology and sericology, 77, 3, 125, 131, 31 Oct. 2008
Japanese Society of Sericultural Science, English

微生物の事典 Ⅲ　　　　　　　　　　　　　　　
朝倉書店, 2008

Construction of the BmNPV T3 Bacmid System and Its Application to the Functional Analysis of BmNPV he65
ONO Chikako, NAKATSUKASA Tomonori, NISHIJIMA Yasuyuki, ASANO Shin-ichiro, SAHARA Ken, BANDO Hisanori, Journal of insect biotechnology and sericology, 76, 3, 161, 167, 30 Oct. 2007
Japanese Society of Sericultural Science, English

Construction of the BmNPVT3 bacmid system and its application to the functional analysis of BmNPV he65
Chikako Ono, Tomonori Nakatsukasa, Yasuyuki Nishijima, Shin Ichiro Asano, Ken Sahara, Hisanori Bando, Journal of Insect Biotechnology and Sericology, 76, 3, 161, 167, 2007

Activation process of the mosquitocidal delta-endotoxin Cry39A produced by Bacillus thuringiensis subsp aizawai BUN1-14 and binding property to Anopheles stephensi BBMV
Takeshi Ito, Hisanori Bando, Shin-ichiro Asano, JOURNAL OF INVERTEBRATE PATHOLOGY, 93, 1, 29, 35, Sep. 2006
English

Cloning and expression of two crystal protein genes, cry30Ba1 and cry44Aa1, obtained from a highly mosquitocidal strain, Bacillus thuringiensis subsp entomocidus INA288
Takeshi Ito, Tomonori Ikeya, Ken Sahara, Hisanori Bando, Shin-ichiro Asano, APPLIED AND ENVIRONMENTAL MICROBIOLOGY, 72, 8, 5673, 5676, Aug. 2006
English

Gene Expression of Autographa californica multiple Nucleopolyhedrovirus (AcMNPV) in Mammalian Cells and Upregulation of Host β-actin Gene
Fujita Ryosuke, Matsuyama Takahiro, Yamagishi Junya, Sahara Ken, Asano Shinichiro, Bando Hisanori, Journal of Virology, 80, 5, 2390, 2395, Mar. 2006
The gene expression of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) was examined in two types of mammalian cells, human HeLa14 and hamster BHK cells. DNA microarray analysis followed by RT-PCR identified at least 12 viral genes transcribed in both HeLa14 cells and BHK cells inoculated with AcMNPV. 5'A RACE was carried out to examine the transcriptional fidelity of these genes in HeLa14 cells. The transcription of ie-1, ie-0 and gp64 was initiated at a baculovirus early gene motif, CAGT, accompanied by a TATA motif. In addition, the same splicing observed for ie-0 mRNA in Sf..., American Society for Microbiology, English

O-05. BAC-FISH analysis of sex-chromosome-autosome translocation in the silkworm(Abstracts of the oral and poster presentations, Abstract of the 56th Annual Meeting of The Society of Chromosome Research)
Ihara Jun-ichiro, Yoshido Atsuo, Asano Shin-ichiro, Bando Hisanori, Sahara Ken, Chromosome science, 9, 1, 12, 12, 2006
The Society of Chromosome Research, English

Marked Decrease of Ribosomal RNA in BmN Cells Infected with AcMNPV
FUJITA Ryousuke, ASANO Shinichiro, SAHARA Ken, BANDO Hisanori, Journal of Insect Biotechnology and Sericology, 74, 3, 125, 128, 28 Oct. 2005
Japanese Society of Sericultural Science, English

Enhancement of cauliflower mosaic virus 35S promoter in insect cells infected with baculovirus
T Abe, N Miyake, Y Nishijima, R Fujita, K Sahara, S Asano, H Bando, VIRUS RESEARCH, 112, 1-2, 38, 41, Sep. 2005
English

家蚕核多角体病ウイルス耐性カイコの作製に関する研究
中務智紀, 小島桂, 田村俊樹, 佐原健, 浅野真一郎, 伴戸久徳, 日本蚕糸学会大会・蚕糸・昆虫機能学術講演会講演要旨集, 75th, 2005

R. Fujita, S. Asano, K. Sahara and H. Bando: Marked decrease of ribosomal RNA in BmN cells infected with AcMNPV, J. Insect Biotech. Seric., 74: 125-128 (2005)*
2005

蚊に強い殺虫活性を有す B. thuringiensis の新規 cry 遺伝子を発見 : 環境に優しく人畜に対して安全な生物農薬を目指して
浅野 眞一郎, 伊藤 岳, 化学と生物, 42, 8, 501, 503, 25 Aug. 2004
日本農芸化学会, Japanese

High frequency Agrobacterium-mediated transformation and plant regeneration via direct shoot formation from leaf explants in Beta vulgaris and Beta maritima
H Hisano, Y Kimoto, H Hayakawa, J Takeichi, T Domae, R Hashimoto, J Abe, S Asano, A Kanazawa, Y Shimamoto, PLANT CELL REPORTS, 22, 12, 910, 918, Jul. 2004
English

Cloning and sequencing of the gene for a putatively nematode-toxic crystal protein, Cry21Ba1, from Bacillus thuringiensis serovar roskildiensis
佐藤 研二, 浅野 真一郎, Japanese journal of nematology, 34, 2, 79, 88, 2004
Bacillus huringiensis serovar roskildiensisから新規なcry遺伝子であるcry21Balをクローニングし、その塩基配列を決定した。本遺伝子のORF（アミノ酸1286残基から成るCry21Balをコードする鎖長3858bpのDNA）の塩基配列は、既知の線虫毒性タンパク質であるCry21Aalをコードする遺伝子（すなわちcry21Aal）のORFの塩基配列と75％一致した。また、Cry21Balの推定アミノ酸配列はCry21Aalのアミノ酸配列と67％一致した。これらの結果は、Cry21Balが線虫毒性タンパク質である可能性を示唆している。, Japanese Nematological Society, English

A strain of Bacillus thuringiensis subsp galleriae containing a novel cry8 gene highly toxic to Anomala cuprea (Coleoptera : Scarabaeidae)
S Asano, C Yamashita, T Iizuka, K Takeuchi, S Yamanaka, D Cerf, T Yamamoto, BIOLOGICAL CONTROL, 28, 2, 191, 196, Oct. 2003
English

Functional Analysis of an Immediate Early Gene, ie1, of Bombyx mori Nucleopolyhedrovirus in Mammalian cells
MATSUYAMA Takahiro, ASANO Shinichiro, SAHARA Ken, BANDO Hisanori, Journal of Insect Biotechnology and Sericology, 72, 2, 87, 94, 30 Jun. 2003
Japanese Society of Sericultural Science, English

Functional Analysis of an Immediate Early Gene, ie1, of Bombyx mori Nucleopolyhedrovirus in Mammalian cells
MATSUYAMA Takahiro, ASANO Shinichiro, SAHARA Ken, BANDO Hisanori, Journal of Insect Biotechnology and Sericology, 72, 2, 87, 94, 30 Jun. 2003
Japanese Society of Sericultural Science, English

W-derived BAC probes as a new tool for identification of the W chromosome and its aberrations in Bombyx mori
Sahara, K., Yoshido, A., Kawamura, N., Ohnuma, A., Abe, H., Mita, K., Oshiki, T., Shimada, T., Asano, S.-I., Bando, H., Yasukochi, Y., Chromosoma, 112, 1, 48, 55, 2003
English

Cloning and Expression of Novel Crystal Protein Genes cry39A and 39orf2 from Bacillus thuringiensis subsp. aizawai Bun1-14 Encoding Mosquitocidal Proteins
ITO Takeshi, SAHARA Ken, BANDO Hisanori, ASANO Shinichiro, Journal of Insect Biotechnology and Sericology, 71, 3, 123, 128, 31 Oct. 2002
Japanese Society of Sericultural Science, English

Antisense and Double-Strand RNA Interference in a Silkworm Ovarian Cell Line
ISOBE Ryoko, KOJIMA Katsura, SAHARA Ken, ASANO Shin-ichiro, BANDO Hisanori, Journal of Insect Biotechnology and Sericology, 71, 1, 43, 47, 28 Feb. 2002
Antisense and double-stranded RNA (dsRNA) interference (RNAi) was investigated in a silkworm ovarian cell line, BmN, using the firefly luciferase as a reporter. The expression of the luciferase from the reporter plasmids was suppressed in the transfected cells when the amount of antisense RNA was in excess of the mRNA amount. The size of the antisense RNA was also found to be an important factor in the interfering efficiency. On the other hand, a small amount of dsRNA induced an efficient RNAi. The sequence-dependency of the dsRNA-derived RNAi was verified in transient expression experiments using the Renillia luciferase, emphasizing the usefulness of dsRNA as a template for gene silencing in the biotechnology of the silkworm., The Japanese Society of Sericultural Science, English

Isobe, R., Kojima, K., Sahara, K., Asano, S. and Bando, H.: "Antisense and double-strand RNA interference in a Silkworm Ovarian Cell Line", J. Insect Biotechnology Sericology, 71:43-47(2002)*　　　　　　　　　　　　　　　
2002

Ito, T., Sahara, K., Bando, H. and Asano, S.: "Cloning and expression of novel crystal protein genes cry39A and 39orf2 from Bacillus thuringiensis subsp. aizawai Bun1-14 encoding mosquitocidal proteins", J. Insect Biotechnology Sericology, 71: 123-128 ・・・
2002
Ito, T., Sahara, K., Bando, H. and Asano, S.: "Cloning and expression of novel crystal protein genes cry39A and 39orf2 from Bacillus thuringiensis subsp. aizawai Bun1-14 encoding mosquitocidal proteins", J. Insect Biotechnology Sericology, 71: 123-128 (2002)*

A simple system for egg production of the wild silkworm, Antheraea yamamai, in large scale
齋藤 寛, 山田 恭裕, 浅野 眞一郎, 伴戸 久徳, 佐原 健, 北海道大学農学部農場研究報告, 32, 0, 71, 74, 29 Mar. 2001
北海道大学農学部附属農場, Japanese

Rearing system for genetic analysis of the wild silkworm, Antheraea yamamai
山田 恭裕, 齋藤 寛, 浅野 眞一郎, 伴戸 久徳, 佐原 健, 北海道大学農学部農場研究報告, 32, 0, 75, 79, 29 Mar. 2001
北海道大学農学部附属農場, Japanese

家蚕核多角体病ウイルスの初期遺伝子発現におけるホルモン応答
小島桂, 浅野真一郎, 佐原健, 伴戸久徳, 東北蚕糸・昆虫利用研究報告, 26, 2001

Tandem repetition of baculovirus ie1 promoter results in upregulation of transcription
K. Kojima, T. Hayakawa, S. Asano, H. Bando, Archives of Virology, 146, 7, 1407, 1414, 2001
Springer Science and Business Media LLC

Comparison of dried-leaf powder of host plants as a component of an artificial diet for the wild silkworm, Antheraea yamamai Guerin-Meneville (Lepidoptera : Saturniidae).
K Sahara, H Saito, K Kikuchi, Y Yamada, S Asano, H Bando, JAPANESE JOURNAL OF APPLIED ENTOMOLOGY AND ZOOLOGY, 45, 2, 94, 98, 2001
Japanese

Analysis of the BmDNV-2 specific DNA polymerase and the common terminal sequence binding proteins
Katsura Kojima, Aya Hirano, Shin-Ichiro Asano, Ken Sahara, Hisanori Bando, Journal of Sericultural Science of Japan, 70, 2, 103, 108, 2001
日本蠶絲學會, English

Comparison of dried-leaf powder of host plants as a component of an artificial diet for the wild silkworm, Antheraea yamamai Guérin-Méneville (Lepidoptera: Saturniidae)
Ken Sahara, Hiroshi Saito, Kunio Kikuchi, Yasuhiro Yamada, Shin-Ichiro Asano, Hisanori Bando, Japanese Journal of Applied Entomology and Zoology, 45, 2, 94, 98, 2001
Chinese

バキュロウイルスの初期遺伝子発現におけるホルモン応答
小島桂, 浅野真一郎, 佐原健, 伴戸久徳, 日本分子生物学会年会プログラム・講演要旨集, 23rd, 2000

PfDNV nonstructural proteins suppress viral RNA splicing
Junya Yamagishi, Shinichiro Asano, Ken Sahara, Toshihiko Iizuka, Hisanori Bando, Journal of Sericultural Science of Japan, 69, 4, 271, 276, 2000
日本蠶絲學會, English

Hayakawa,T., Kojima, K., Nonaka, K., Nakagaki, M., Sahara, K., Asano, S., Iizuka, T. and Bando, H. : Analysis of protein encoded in the bipartite genome of a new type of parvo-like virus isolated from silkworm -structural protein with DNA polymerase mo・・・　　　　　　　　　　　　　　　
2000
Hayakawa,T., Kojima, K., Nonaka, K., Nakagaki, M., Sahara, K., Asano, S., Iizuka, T. and Bando, H. :

Analysis of protein encoded in the bipartite genome of a new type of parvo-like virus isolated from silkworm -structural protein with DNA polymerase motif. Virus Research 66, 101-108 (2000)

PfDNV nonstructural proteins suppress viral RNA splicing
Junya Yamagishi, Shinichiro Asano, Ken Sahara, Toshihiko Iizuka, Hisanori Bando, Journal of Sericultural Science of Japan, 69, 4, 271, 276, 2000
English

カイコ培養細胞を用いた持続的発現系の開発 プロモーターの改良 その2
小島桂, 浅野真一郎, 佐原健, 飯塚敏彦, 伴戸久徳, 東北蚕糸研究報告, 24, 1999

Toxicity of Bacillus thuringiensis subsp. wuhanensis crystal protein to Bombyx mori and Spodoptera litura
Yulia Pujiastuti, Shin-ichiro Asano, Ken Sahara, Hlsanori Bando, Toshihiko Ilzuka, Journal of Sericultural Science of Japan, 68, 3, 195, 199, 1999
日本蠶絲學會, English

Yulia Pujiastuti, Shin-ichiro Asano, Ken Sahara, Hisanori Bando and Toshihiko Iizuka : "Toxicity of Bacillus thuringiensis subsp. wuhanensis crystal protein to Bombyx mori and Spodoptera litura", J. Seric. Sci. Jpn 68(3), 195-200, (1999)　　　　　　　　　　　　　　　
1999

New Japanese isolates of Bacillus popilliae isolated from milky diseased larvae of Popillia japonica, Anomala rufocuprea and Anomala daimiana (Coleoptera : Scarabaeidae) (vol 32, pg 583, 1997)
N Matsuki, S Asano, H Bando, T Iizuka, APPLIED ENTOMOLOGY AND ZOOLOGY, 33, 1, 205, 205, Feb. 1998
English, Others

Novel Bacillus thuringiensis serovar aizawai strains isolated from mulberry leaves in Indonesia
A Rizali, S Asano, K Sahara, H Bando, BW Lay, S Hastowo, T Iizuka, APPLIED ENTOMOLOGY AND ZOOLOGY, 33, 1, 111, 114, Feb. 1998
English

家蚕核多角体病ウイルス遺伝子プロモーターの改良とその応用
小島桂, 浅野真一郎, 佐原健, 飯塚敏彦, 伴戸久徳, 東北蚕糸研究報告, 23, 1998

Identification of cryl genes from Bacillus thuringiensis strains which have activity against Spodoptera litura.
Shin-ichiro Asano, Yulia Pujiastuti, Ken Sahara, Hisanori Bando, Toshihiko Iizuka, Har unori Kikuta, Journal of Sericultural Science of Japan, 67, 3, 237, 242, 1998
日本蚕糸学会, English

Identification of cry1 genes from Bacillus thuringiensis strains which have activity against Spodoptera litura.
ASANO SHIN-ICHIRO, PUJIASTUTI YULIA, SAHARA KEN, BANDO HISANORI, KIKUTA HARUNORI, IIZUKA TOSHIHIKO, Journal of Insect Biotechnology and Sericology, 67, 3, 237, 242, 1998
Bacillus thuringiensis reference strains and isolates used in this experiment were stored in our laboratory and originally isolated from soil and dead insects in Hokkaido. In order to find B. thuringiensis strains which have a high toxic activity against Spodoptera litura larvae, these B. thuringiensis strains were bioassayed to the 3rd-instar larvae of S. litura.
The identification of cry1 genes from active strains against S. litura larvae were demonstrated by using PCR. Cry1C cloned from serovar entomocidus and serovar kenyae revealed a toxic activity against S. litura larvae. On the contrary, cry1E gene cloned from serovar tolworthi and serovar darmstadiensis did not have a toxic activity.
On the other hand, even though serovar galleriae Acp10-8 and wuhanensis have a highly toxic activity against S. litura larvae, both of strains did not involve cry1C and cry1E genes. It seems that they have the novel cry genes., The Japanese Society of Sericultural Science, Japanese

New Japanese isolates of Bacillus popilliae isolated from milky diseased larvae of Popillia japonica, Anomala rufocuprea and Anomala daimiana (Coleoptera: Scarabaeidae)
N Matsuki, S Asano, H Bando, T Iizuka, APPLIED ENTOMOLOGY AND ZOOLOGY, 32, 4, 583, 588, Nov. 1997
English

New Japanese isolates of Bacillus popilliae isolated from milky diseased larvae of Popillia japonica, Anomala rufocuprea and Anomala daimiana (Coleoptera: Scarabaeidae)
N Matsuki, S Asano, H Bando, T Iizuka, APPLIED ENTOMOLOGY AND ZOOLOGY, 32, 4, 583, 588, Nov. 1997
English

Isolation and Insecticidal Crystal Protein Gene Screening of Bacillus thuringiensis from Kirisima Yakushima Island national park
KIKUTA Harunori, KUROIWA Manabu, ASANO Shinichiro, IIZUKA Toshihiko, Journal of the College of Dairying. Natural science, 22, 1, 81, 97, Oct. 1997
Rakuno Gakuen University, Japanese

Hatchability in triploid silkworms of sex-linked chocolate
SAHARA KEN, FUKUTANI SHOUKO, SAITOH HIROSHI, NAKADA TOHRU, ASANO SHIN-ICHIRO, BANDO HISANORI, KAWAMURA NAOKO, IIZUKA TOSHIHIKO, The Journal of Sericultural Science of Japan, 66, 2, 141, 144, 28 Apr. 1997
The Japanese Society of Sericultural Science, Japanese

Effective length of starvation for acquiring fertile tetrtaploid males in the silkworm
Ken Sahara, Youko Tanaka, Yasuhiro Yamada, Hiroshi Saitoh, Tohru Nakada, Shin-ichiro Asano, Hisanori Bando, Toshihiko Iizuka, Naoko Kawamura, Journal of Sericultural Science of Japan, 66, 3, 207, 211, 1997
日本蚕糸学会, English

tetraploid silkworms from tetraploid parents
Ken Sahara, Yasuhiro Yamada, Hiroshi Saitoh, Tohru Nakada, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Naoko Kawamura, Journal of Sericultural Science of Japan, 66, 5, 341, 345, 1997
日本蠶絲學會, English

Egg size of knobbed silkworm
Ken Saharan, P. Rama, Yasuhiro Yamada, Hiroshi Saitoh, Shin-Ichiro Asano, Hisanori Bando, Toshihiko Iizuka, Tohru Nakada, Mohana Rao, Journal of Sericultural Science of Japan, 66, 5, 364, 366, 1997
日本蚕糸学会, English

Detection of the silkworm-pathogenic virus genomes by PCR.
KAGEYASU SEIJI, HAYAKAWA TOHORU, ISAWA HARUHIKO, ASANO SHINICHIRO, SAHARA KEN, IIZUKA TOSHIHIKO, BANDO HISANORI, The Journal of Sericultural Science of Japan, 66, 6, 477, 483, 1997
The five viruses, Bombyx mori densonucleosis virus type 1 (BmDNV-1), -type 2 (BmDNV-2), Bombyx mori nuclear polyhedrosis virus (BmDPV), Bombyx mori cytoplasmic polyhedrosis virus (BmCPV), and Infectious flacherie virus (IFV), are the agents causing fetal disease of the silkworm. The aim of this study is to establish a simple and highly sensitive PCR system for detection of all the viruses. A combination of the RT-PCR and the nested PCR was found to be effective to amplify DNA and/or RNA virus genome sequences in a reaction maintaining high sensitivity and specificity. In this report, the primer design and the condition of the PCR for the detection of all the viruses are described., The Japanese Society of Sericultural Science, Japanese

Iizuka, T., Yamaya, K., Rizali, A., Asano, S. -i., Bando, H., Hastowo, S. and Lay, B. W. : "Isolation and identification of a new mosquitocidal isolate, Bacillus thuringiensis subsp. entomocidus INA288", (T. Attathom et al. : Bacillus thuringiensis Bio・・・　　　　　　　　　　　　　　　
1997
Iizuka, T., Yamaya, K., Rizali, A., Asano, S. -i., Bando, H., Hastowo, S. and Lay, B. W. : "Isolation and identification of a new mosquitocidal isolate, Bacillus thuringiensis subsp. entomocidus INA288", (T. Attathom et al. : Bacillus thuringiensis Biotechnology and Environmental Benefits, 2 : 152-160 (1997)＊

Effective length of starvation for acquiring fertile tetrtaploid males in the silkworm
Ken Sahara, Youko Tanaka, Yasuhiro Yamada, Hiroshi Saitoh, Tohru Nakada, Shin-ichiro Asano, Hisanori Bando, Toshihiko Iizuka, Naoko Kawamura, Journal of Sericultural Science of Japan, 66, 3, 207, 211, 1997
English

Hatchability in triploid silkworms of sex-linked chocolate
SAHARA KEN, FUKUTANI SHOUKO, SAITOH HIROSHI, NAKADA TOHRU, ASANO SHIN-ICHIRO, BANDO HISANORI, KAWAMURA NAOKO, IIZUKA TOSHIHIKO, Journal of Insect Biotechnology and Sericology, 66, 2, 141, 144, 1997
The Japanese Society of Sericultural Science, Japanese

Detection of the silkworm-pathogenic virus genomes by PCR.
KAGEYASU SEIJI, HAYAKAWA TOHORU, ISAWA HARUHIKO, ASANO SHINICHIRO, SAHARA KEN, IIZUKA TOSHIHIKO, BANDO HISANORI, The Journal of Sericultural Science of Japan, 66, 6, 477, 483, 1997
The five viruses, Bombyx mori densonucleosis virus type 1 (BmDNV-1), -type 2 (BmDNV-2), Bombyx mori nuclear polyhedrosis virus (BmDPV), Bombyx mori cytoplasmic polyhedrosis virus (BmCPV), and Infectious flacherie virus (IFV), are the agents causing fetal disease of the silkworm. The aim of this study is to establish a simple and highly sensitive PCR system for detection of all the viruses. A combination of the RT-PCR and the nested PCR was found to be effective to amplify DNA and/or RNA virus genome sequences in a reaction maintaining high sensitivity and specificity. In this report, the primer design and the condition of the PCR for the detection of all the viruses are described., The Japanese Society of Sericultural Science, Japanese

Hayakawa, T., Asano, S. -i., Sahara, K., Iizuka, T. and Bando H. : "Detection of replicative intermediate with closed terminus of Bombyx densonucleosis virus", Arch. Virol., 142 : 1-7 (1997)＊　　　　　　　　　　　　　　　
1997

tetraploid silkworms from tetraploid parents
Ken Sahara, Yasuhiro Yamada, Hiroshi Saitoh, Tohru Nakada, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Naoko Kawamura, Journal of Sericultural Science of Japan, 66, 5, 341, 345, 1997
English

Asano, S. -i., Nukumizu, Y., Bando, H., Iizuka, T. and Yamamoto, T. : "Cloning of novel enterotoxin genes from Bacillus cereus and Bacillus thuringiensis", Appl. Environ. Microbiol., 63(3) : 1054-1057 (1997)＊　　　　　　　　　　　　　　　
1997

Asano, S. -i., Okumura, E., Kamagata, Y., Sahara, K., Bando, H., Iizuka, T. and Yamamoto, T. : "Mutagenesis of loop3 region of the Bacillus thuringiensis Cry1Ac delta-endotoxin affect DBM's receptor binding", (T. Attathom et al. : Bacillus thuringiensi・・・　　　　　　　　　　　　　　　
1997
Asano, S. -i., Okumura, E., Kamagata, Y., Sahara, K., Bando, H., Iizuka, T. and Yamamoto, T. : "Mutagenesis of loop3 region of the Bacillus thuringiensis Cry1Ac delta-endotoxin affect DBM's receptor binding", (T. Attathom et al. : Bacillus thuringiensis Biotechnology and Environmental Benefits, 2 : 49-60 (1997)＊

Sasaki, J., Asano, S. -i., Hashimoto, N., Lay, B. W., Hastowo, S., Bando, H. and Iizuka, T. : "Characterization of a cry2A gene cloned from an isolate of Bacillus thuringiensis serovar sotto", Curr. Microbiol., 35 : 1-8 (1997)＊　　　　　　　　　　　　　　　
1997

Egg size of knobbed silkworm
Ken Saharan, P. Rama, Yasuhiro Yamada, Hiroshi Saitoh, Shin-Ichiro Asano, Hisanori Bando, Toshihiko Iizuka, Tohru Nakada, Mohana Rao, Journal of Sericultural Science of Japan, 66, 5, 364, 366, 1997
English

Insecticidal activity of the protein encoded by the cryV gene of Bacillus thuringiensis kurstaki INA-02
Jun Sasaki, Shinichiro Asano, Toshihiko Iizuka, Hisanori Bando, Bibiana W. Lay, Sugyo Hastowo, Gary K. Powell, Takashi Yamamoto, Current Microbiology, 32, 4, 195, 200, Apr. 1996
English

蚊に殺虫活性を有する新規Bacillus thuringiensis subsp.entomocidus INA 288の分離と性状 : 微生物が生産する殺虫性蛋白質 : グラム陽性菌Bacillus thuringiensisとその利用 : シンポジウム(S-2)
飯塚 敏彦, 山谷 公美子, Rizali Akhmad, 浅野 真一郎, 伴戸 久徳, Hastowo Sugyo, Lay Bibiana W, 日本農藝化學會誌, 70, 0, 339, 339, 05 Mar. 1996
社団法人日本農芸化学会, Japanese

Identification of cry Gene from Bacillus thuringiensis by PCR and isolation of unique insecticidal bacteria
Asano Shin-ichiro, Memoirs of the Faculty of Agriculture, Hokkaido University, 19, 7, 529, 563, 1996
Hokkaido University, Japanese

Identification of cry V gene from Bacillus thuringiensis isolates
Jun Sasaki, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Bibiana W. Lay, Sugyo Hastowo, Journal of Sericultural Science of Japan, 65, 1, 56, 61, 1996
日本蚕糸学会, English

Bacillus thuringiensis ICP gene expression under the control of cry I A (a) gene promoter.
Naoki Hashimoto, Jun Sasaki, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Journal of Sericultural Science of Japan, 65, 3, 185, 191, 1996
日本蚕糸学会, English

Bacillus thuringiensis ICP gene expression under the control of cry I A (a) gene promoter.
Naoki Hashimoto, Jun Sasaki, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Journal of Sericultural Science of Japan, 65, 3, 185, 191, 1996
English

Identification of cry V gene from Bacillus thuringiensis isolates
Jun Sasaki, Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Bibiana W. Lay, Sugyo Hastowo, Journal of Sericultural Science of Japan, 65, 1, 56, 61, 1996
English

「ＰＣＲ法による Bacillus thuringiensis cry 遺伝子同定法の確立と新規 B. thuringiensis 菌株の発見」　　　　　　　　　　　　　　　
『北大農邦文紀要』, 19, 7, 529, 563, 1996

Enhancing Effects of Supernatants from Various Cultures of Bacillus thuringiensis on Larvicidal Activity of δ-Endotoxin against the Common Cutworm, Spodoptera litura
ASANO Shoji, HORI Hidetaka, Applied entomology and zoology, 30, 2, 369, 374, 1995
The synergistic effects of supernatants from cultures of nine strains of Bacillus thuringiensis toward the insecticidal activity of δ-endotoxin were investigated using neonates of the common cutworm, Spodoptera litura, and late 3rd instar larvae of the diamondback moth, Plutella xylostella by the diet incorporation method. Synergistic effects between supernatants and δ-endotoxins were detected against S. litura but not toward P. xylostella. Synergistic activity in supernatants against S. litura but not toward P. xylostella. Synergistic activity in supernatants against S. litura was observed in four strains and the occurrence was shown to be dependent on culture media. Interestingly, B. thuringiensis serovar japonensis strain Buibui and B. thuringinensis serovar morrisoni strain san diego, which produce insecticidal crystal proteins specific only to coleopteran larvae, were shown to produce synergistic factor(s) in culture supernatants. The synergistic substance(s) released into supernatants from various strains of B. thuringiensis were apparently the same., Japanese Society of Applied Entomology and Zoology, English

AMPLIFICATION AND DETECTION OF CRYIIIA GENE FROM BACILLUS-THURINGIENSIS STRAINS
S ASANO, J SASAKI, H BANDO, T IIZUKA, JAPANESE JOURNAL OF APPLIED ENTOMOLOGY AND ZOOLOGY, 38, 4, 300, 302, Nov. 1994
Japanese, Introduction scientific journal

Bacillus thuringiensis subsp. israelensis の cryIVC および cryIVD 遺伝子の検索
浅野真一郎, 東北蚕糸研究報告, 19, 34, 34, 1994

E320 双翅目昆虫に活性のあるB.thuringiensis殺虫性タンパク質遺伝子(CryIV)の増幅と同定(生物的防除・微生物防除)
浅野 真一郎, 佐々木 潤, 伴戸 久徳, 飯塚 敏彦, 菊田 治典, 日本応用動物昆虫学会大会講演要旨, 0, 37, 146, 146, 03 Apr. 1993
日本応用動物昆虫学会, Japanese

E325 Bacillus cereusのエンテロトキシン遺伝子のクローニングと一部構造解析(生物的防除・微生物防除)
温水 友紀, 浅野 真一郎, 伴戸 久徳, 飯塚 敏彦, 日本応用動物昆虫学会大会講演要旨, 0, 37, 149, 149, 03 Apr. 1993
日本応用動物昆虫学会, Japanese

E326 北海道のゴルフ場において罹病コガネムシ類から分離されたBacillus popilliaeの性状(生物的防除・微生物防除)
松木 伸浩, 浅野 真一郎, 伴戸 久徳, 飯塚 敏彦, 日本応用動物昆虫学会大会講演要旨, 0, 37, 149, 149, 03 Apr. 1993
日本応用動物昆虫学会, Japanese

Amplification and identification of cry ii genes from bacillus thuringiensis by PCR procedure
Shinichiro Asano, Hisanori Bando, Toshihiko Iizuka, Journal of Sericultural Science of Japan, 62, 3, 223, 227, 1993
日本蚕糸学会, English

Identification of cry II genes in bacillus thuringiensis isolates by the use of oligo-nucleotide DNA primers
Shinichirou Asano, Hisanori Bando, Toshihiko Iizuka, Harunori Kikuta, Journal of Sericultural Science of Japan, 62, 3, 210, 215, 1993
日本蠶絲學會, English

E215 鞘翅目昆虫に活性のあるB. thuringiensis殺虫性タンパク質遺伝子のクローニングと発現タンパク質の殺虫活性(微生物防除)
浅野 真一郎, 佐々木 潤, 伴戸 久徳, 飯塚 敏彦, 日本応用動物昆虫学会大会講演要旨, 0, 36, 211, 211, 10 Sep. 1992
日本応用動物昆虫学会, Japanese

E216 Cry I, Cry II両遺伝子を有するBacillus thuringiensisのヤマトヤブカに対する殺虫活性(微生物防除)
菊田 治典, 浅野 眞一郎, 伴戸 久徳, 飯塚 敏彦, 日本応用動物昆虫学会大会講演要旨, 0, 36, 212, 212, 10 Sep. 1992
日本応用動物昆虫学会, Japanese

Identification of insecticidal activity of Bacillus thuringiensis against the silkworm by DNA hybridization method.
ASANO SHINICHIRO, IIZUKA TOSHIHIKO, Journal of Insect Biotechnology and Sericology, 60, 6, p475, 479, Dec. 1991
In order to identify the insecticidal activity of 20 Bacillus thuringiensis subspecies which produce bipyramidal crystals, dot blot hybridization of DNA purified from the subspecies was performed with a cloned crystal protein gene (cryl-1 gene) from subsp. kurstaki HD-1. The DNAs from the ten subspecies exhibiting on insecticidal activity against the silkworm, Bombyx mori, strongly hybridized with the probe prepared from the cloned crystal protein gene. However, DNAs from the other ten subspecies lacking insecticidal activity did not hybridize with the probe. Similar results were also obtained in the experiment using a probe prepared from the DNA fragment that is specific for insecticidal activity. Based on these results, we concluded that the intensity of the insecticidal activity against the silkworm can be determined DNA hybridization., The Japanese Society of Sericultural Science, Japanese

B37 数種プロテアーゼインヒビターの家蚕に対する影響(昆虫病理・微生物防除)
飯塚 敏彦, 浅野 真一郎, 日本応用動物昆虫学会大会講演要旨, 0, 35, 103, 103, 15 Sep. 1991
日本応用動物昆虫学会, Japanese

Protoxin from diamond-shaped crystals from B. thuringiensis and insecticidal activity against the silkworm, Bombyx mori.
ASANO SHINICHIRO, IIZUKA TOSHIHIKO, Journal of Insect Biotechnology and Sericology, 59, 5, 375, 380, Oct. 1990
The amount of protoxin from the diamond-shaped clystals produced by twenty Bacillus thuringiensis subspecies/strains was compared with the peptide level using SDS treatment as a simple and improved method. The insecticidal activities of the spore and crystal samples from these twenty strains were tested against 5th instar larvae of the silkworm by peroral injection. Lepidopteran insecticidal activities were recognized in the strains in which 130 to 140kDa peptides were detected except for the subsp. japonensis in which 120kDa protoxin was not detected. However, insecticidal activity was not revealed in the strains in which 130 to 140kDa peptides were not detected., The Japanese Society of Sericultural Science, Japanese

Purification of crystals from Bacillus thuringiensis by using Percoll
Fujio Baba, Shinichiro Asano, Toshihiko Iizuka, Journal of Sericultural Science of Japan, 59, 6, 487, 489, 1990
日本蚕糸学会, English

Isolates of Bacillus thuringiensis from soil at Hokkaido.
KIKUTA H, Memoirs of the Faculty of Agriculture,Hokkaido University, 16, 4, p383, 389,図8p, 1989
北海道大学, Japanese

バイオロジカル・コントロール－害虫管理と天敵の生物学－　　　　　　　　　　　　　　　
朝倉書店, 2009

Evaluation of Biological Control of Oryctes Rhinoceros beetle contributes to the protection of palms in the Pacific Ocean
Grants-in-Aid for Scientific Research
07 Oct. 2021 - 31 Mar. 2025
仲井 まどか, 浅野 眞一郎, 坂本 卓磨
タイワンカブトムシ(別名サイカブトムシ：Oryctes rhinoceros）は、ヤシ類の害虫である。本種の防除には、1970年代から1980年代にかけて天敵ウイルス（Oryctes rhinoceros nudivirus: OrNV) を用いた防除が実施され、南太平洋のフィジーなどではヤシの被害を抑えることに成功した。しかし、2007年にグアム（米国）にウイルス抵抗性のハプロタイプ（Gタイプ）が侵入したことが報告され、その後もGタイプの侵入が、パプアニューギニア、ソロモン諸島、ハワイなどに拡大していることから太平洋州の各国でヤシ類の植物保護において大きな脅威になっている。また、パラオにはGタイプとそれ以外のハプロタイプ（Sタイプ）が混在しておりヤシの被害も少ない。本研究は、Gタイプの害虫としての特徴と天敵ウイルスを用いた生物的防除の機構を解明することによりその防除の成否を決定する要因の解明を目的とする。本研究において国際共同研究を強化することにより、太平洋州全域のヤシ林の保護を実現させることを目指している。
2021年度は、新型コロナウイルス感染拡大のため、GタイプおよびSタイプの輸入ができなかった。そこで、天敵ウイルスによる本種の防除メカニズムの解明について研究を行った。具体的には、幼虫および成虫に対するウイルス接種法の検討を行った。接種方法が確立できれば、宿主のステージごとのウイルス伝播率を定量化することができる。室内実験においてウイルスの伝播率が、幼虫から幼虫、幼虫から成虫、成虫から幼虫、成虫から成虫のどのステージごとにどれだけ伝播するか比較できれば、 OrNVを用いた生物的防除の成否を決定するメカニズムの解明に繋がる。また、培養細胞を用いたウイルス増殖を検討した。さらに、沖縄県において採集したタイワンカブトムシを大量飼育し、生物検定の基盤確立を行った。
Japan Society for the Promotion of Science, Fund for the Promotion of Joint International Research (Fostering Joint International Research (B)), Tokyo University of Agriculture and Technology, 21KK0111

Factors associated with occurrence of novel biotype of Oryctes rhinoceros resistant against nudiviruses
Grants-in-Aid for Scientific Research
01 Apr. 2017 - 31 Mar. 2021
Nakai Madoka
Coconut Rhinoceros Beetle (Oryctes rhinoceros) is a pest of coconut palms and oil palms. Natural enemy of this species, nudivirus, was used to control this species in the 1970s and 1980s, and succeeded in reducing damage to palms in the South Pacific including Fiji. However, in 2007, it was reported that a haplotype (a strain of the same species with a different genotype), in which the virus is ineffective, invaded Guam (USA), and since then this G-type has been expanding its distribution to Palau, Hawaii, Solomon Islands, Papua New Guinea, and other countries. In this study, we isolated natural enemy viruses from Palau populations in order to search for virus strains that could be used to control the G-type.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Tokyo University of Agriculture and Technology, 17H04622

Functional genomics on in vivo fitness of BmNPV
Grants-in-Aid for Scientific Research
01 Apr. 2015 - 31 Mar. 2017
Hisanori Bando, ASANO SHINICHIRO
Bombyx mori nucleopolyhedrovirus (BmNPV) H4 that had genome sharing high sequence similarity (approx. 97%) with that of the type strain of BmNPV T3 showed marked growth advantage in silkworm larvae. Studies using chimeric recombinant BmNPV between H4 and T3 demonstrated that a viral membrane protein gene gp64 as a determinant in the in vivo fitness of BmNPV H4. Further analyses identified amino acid positions involved in the BmNPV H4 phenotype of efficient growth in silkworm larvae. These results suggested the possibility to control growth ability of BmNPV in silkworm larvae by gp64-targetted genome manipulation.
Japan Society for the Promotion of Science, Grant-in-Aid for Challenging Exploratory Research, Hokkaido University, 15K14891

Elucidation of insecticidal activity mechanism of B. thuringiensis Cry44Aa toxin
Grants-in-Aid for Scientific Research
2014 - 2016
ASANO SHINICHIRO
I have been conducting research aiming at elucidating the insecticidal activity mechanism of Cry44Aa toxin produced by Bacillus thuringiensis entomocidus INA 288, which is known to have highly insecticidal activity against Aedes aegypti and Culex pippins which intermediate tropical infectious diseases. As a result of research for Cry44Aa toxin receptor molecules in A. aegypti which play an important role in insecticidal activity mechanism, alkaline phosphatase and ABC transferase were identified as candidate molecules.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, Principal investigator, Competitive research funding, 26450463

Mechanisms of Dual Control using Entomopathogenic Fungi and Bacteria
Grants-in-Aid for Scientific Research
01 Apr. 2012 - 31 Mar. 2015
MASNAORI Koike, AIUCHI Daigo, ASANO Shin-ichiro, MASUDA Toshio, SEKINE Muneyuki
Fungal entomopathogens have been widely investigated as biological control agents of pest insects in attempts to improve the sustainability of crop protection. Simultaneous biological control of both insect pests and plant pathogens (dual control) has been reported for the hypocrealean fungal entomopathogens, Beauveria bassiana, Metharizium spp. and Lecanicillium spp. And accumulating evidence shows that Beauveria spp. can colonize a wide array of plant species endophytically. Furthermore, traits that are important for insect pathogenicity are also involved in pathogenicity to plant pathogen and plant pathogenic nematode.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), Obihiro University of Agriculture and Veterinary Medicine, 24380174

Development of new BT agents and Coleoptera adult insecticidal activity mechanism
Grants-in-Aid for Scientific Research
2010 - 2012
ASANO Shinichiro
To find the novel Cry toxin with specific insecticidal activity in adult Coleoptera pests, we were screening by PCR and Bioassay from our B. thuringiensis strains’ library. As microbial control agent against Coleoptera adults to conduct for the scarab beetle adult and leaf beetles adult of Cry8D toxin, and to elucidate the insecticidal activity mechanism adult-specific, is a flame pest control, I develop a new BT agents.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (C), Hokkaido University, Principal investigator, Competitive research funding, 22580056

病原微生物を用いた害虫防除　　　　　　　　　　　　　　　
科学研究費補助金
2008 - 2012
Competitive research funding

Microbiol Pest Management　　　　　　　　　　　　　　　
Grant-in-Aid for Scientific Research
2008 - 2012
Competitive research funding

殺虫性結晶タンパク質産生菌の分子生物学的研究　　　　　　　　　　　　　　　
共同研究
2008
Competitive research funding

Research for molecilar biology of Bacillus thuringiensis insecticidal crystal protein　　　　　　　　　　　　　　　
Cooperative Research
2008
Competitive research funding

Molecular Basis for Safety Assessment of Microbial Pesticides
Grants-in-Aid for Scientific Research
2003 - 2006
BANDO Hisanori, ASANO Shinichiro, SAHARA Ken
Baculovirus and Bt (Bacillus thuringiensis) are used as microbial pesticides all over the world and the former also provide a most efficient foreign gene expression system in eukaryotes (baculovector system). We studied the molecular basis in the safety of these entomopathogenic microorganisms. Recent studies demonstrated that a baculovirus (AcMNPV) can enter the nucleus of mammalian cells independently of the cell cycle without multiplication and effectively express foreign genes without expressing its own genes, suggesting that the baculovirus can be used as a low risk viral vector in the field of gene therapy. Contrary to the general perception, we elucidated that AcMNPV expressed viral genes at least partly in mammalian cells through the usual infection pathway. In addition, the expression of cellular gene (at least β-actin) was upregulated in the mammalian cells inoculated with AcMNPV. However, the expression of viral genes was inhibited to low level in mammalian cells partly by histone deacetylation mechanism and further the transcription of some viral genes started at incorrect site(s).
As for the B. thuringiensis (Bt) strains, we investigated mainly the mechanisms of Vero cell toxicity by non-hemolytic enterotoxin C (NheC) that was thought to be the most important enterotoxin causing food-poisoning. It was elucidated that NheC secreted from a clinical isolate of B. cereus (Bc) strain possessing Vero cell toxicity could attach to the surface of Vero cells through the N-terminal region of 30 amino acids. On the other hand, the corresponding N-terminal region was truncated possibly by post-translational cleavage in the NheC secreted from Bt strains used in the commercial products. In addition, the N-terminal truncated from of NheC didn't show Vero cell toxicity any more.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), Hokkaido University, 15208006

A New approach to the practical use of insect-tolerant lines of sugar beet via foreign gene transfer
Grants-in-Aid for Scientific Research
1999 - 2002
KANAZAWA Akira, IKEGUCHI Shojiro, ASANO Shin-ichiro
The larvae of cabbage armyworm (Mamestra brassicae L.) bring heavy damage on the final yield of beet-roots and subsequently, sugar production, by eating stems and leaves of sugar beet plants. In order to establish sugar beet cultivars that are tolerant to insects including cabbage armyworm, we have been developing transgenic sugar beet lines harboring cryIA(b), a gene encoding insecticide protein from Bacillus thuringiensis. In the present study, based on the results so far obtained, we tried to make transgenic sugar beet lines that show increased insecticide activity aiming at practical use of the transgenic lines. First, we introduced cryIC into sugar beet plants and analyzed the effect of its expression. We chose cryIC from various candidates of insecticide protein genes, since its gene product showed highest insecticide activity against cabbage armyworm among various insecticide proteins that were obtained by expressing cloned genes in E. coli. Meanwhile, reports were published that described that the introduction of insecticide protein genes into chloroplast gave increased accumulation of the protein and consequently, gave increased insecticide activity of plants. We, therefore, developed vectors available for chloroplast transformation and selection system of transformed chloroplast in order to introduce insecticide protein genes into sugar beet chloroplasts. We also succeeded in introducing several genes practically useful for sugar beet production and evaluated the effect of gene expression. In addition, we analyzed the stability of the expression of foreign genes in plant cells in order to make sure their experssion.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), HOKKAIDO UNIVERSITY, 11356001

Analysis of novel insecticidal crystal protein genes from B.thuringiensis and application of biological insecticide
Grants-in-Aid for Scientific Research(基盤研究(B))
1999 - 2000
飯塚 敏彦, Shin-ichiro ASANO, 浅野 眞一郎, Ken SAHARA
1. Screening of a scarab-cidal B.thuringiensis (1999)In order to find of a scarab-cidal B.thuringiensis, we were screening our B.thuringiensis stocks.2. Analysis of the scarab-cidal B.thuringiensis strain (gallariae BBT2-8)(2000)We found the scarab-cidal Bt.BBT2-8, then we cloned the crystal protein genes (cryB28 gene) have a scarab-cidal activity. This cryB28 gene was expressed in E.coli and acrystariferous B.thuringiensis strain (BT51). The expression products had a highly activity against scarabe.3. Screening of a nematoda-cidal B.thuringiensis (1999-2000)In order to find of a nematoda-cidal B.thuringiensis, we were screening our B.thuringiensis stocks by using PCR.We could not find nematodacidal B.thuringiensis strains.4. Screening of novel lepidopteran-cidal B.thuringiensis (1999)In order to find of novel lepidopteran-cidal B.thuringiensis, we were screening our B.thuringiensis stocks. We picked up the novel lepidopteran acitive Bt strain (wuhanensis), then we cloned the novel cry1 gene (cry1Db1) from wuhanensis have insecticidal activities against Spodoptera litura, Plutella xylostella and Bombyx mori.
Ministry of Education, Culture, Sports, Science and Technology, 基盤研究(B), 北海道大学, Principal investigator, Competitive research funding, 11460026

コガネムシに殺虫活性を有するB.popilliaeの結晶タンパク質遺伝子解析
科学研究費補助金(萌芽的研究)
1998 - 1999
浅野 眞一郎
当該研究者は既に、北海道のゴルフ場にて数種のコガネムシを採取して、そこからB.popilliae菌株(3菌株、var.Mame,var.Hime,var.Sakura)を分離し、それらのB.popilliae菌株の結晶タンバク質を解析した。各菌株の結晶タンパク質のSDS-PAGEによる解析の結果、それぞれの菌株が約80kDaのペプチドからなる結晶タンパク質を産生していることが明らかとなった。そこで、var.Mame株の80kDaのペプチドのN末端アミノ酸配列の解析(10残基)を行った。その結果、報告されている、B.popilliae由来のタンパク質との相同性は認められなかった。既に報告されている、B.popilliae由来の結晶タンパク質遺伝子(cryBP1:cry18A)に特異なオリゴヌクレオチドプライマーを合成し、B.popilliae菌株のcry遺伝子の検索をPCR法を用いて行った。各種プライマーを用いてcry遺伝子検索を行った結果、これらのB.popilliae菌株が、報告されているcryBP1遺伝子を有しないことが明らかとなった。これらのB.popilliae菌株の有するcry遺伝子をクローニングするため、cryBP1遺伝子の上流域に存在するorf1に注目し、このorf1の塩基配列を基にDNAプライマーを作成した。そのプライマーを用いて、B.popilliae var popilliae株のゲノムDNAをテンプレートとして、PCR法によって約300bpの断片が増幅された。この増幅された断片を解析したところ、orf1と高い相同性が認められた。現在は、この増幅されたDNA断片の上・下流域のクローニングを進めている。
文部科学省, 萌芽的研究, 北海道大学, Principal investigator, Competitive research funding, 10876010

Studies on Development of Efficient Transgenic System for Alien Gene of Insecticidal Protein in Sugarbeet
Grants-in-Aid for Scientific Research
1997 - 1999
SHIMAMOTO Yoshiya, KANAZAWA Akira, ASANO Shinichiro, SANO Yoshio
1)Development of chloroplast transgenic system in sugarbeet
We have progressed to develop a chloroplast transgenic system in sugarbeet in which is introduced cry genes to chloroplast genome and produce efficiently insecticidal crystal protein (ICP) in chloroplast. Plasmid carrying selective factor (aadA), marker factor (GUS), promoter (rrn) and terminator (rps 16) was incorporated into disc tissue of shoot of sugarbeet by microprojectile bombardment. In the tissue culture regeneration plant has not achieved. Repeatedly plasmid was constructed and incorporated into the sugarbeet.
2)Development of concentration system of protein upon organelle in sugarbeet
We have progressed to develop a concentration system of toxic protein upon organelle in sugarbeet. Base sequence coding transit peptide to chloroplast was cloned from tobacco plant and was linked with position of N terminal in GUS gene. Thus the plasmid was constructed and introduced into sugarbeet by Agrobacterium method.
3)Expression of cry gene in transformant sugarbeet
Several transformants carrying cryIA(b) were detected by PCR southern, ICP analysis and PCR southern blotting in half of plants regenerating after procedure of Agrobacterium method. Several leaves of these transformants were supplied for cabbage armyworm with the third larva age. Larva of cabbage armyworm was able to grow normally on leaves with non-cry gene and not to grow on several leaves of transformant.
4)Screening for efficient cry gene to produce insecticidal protein
Screening for efficient cry gene to produce insecticidal protein was conducted for cabbage armyworm. CryIC gene has more insecticidal ability than cryIA(b) in larva of cabbage armyworm.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), HOKKAIDO UNIVERSITY, 09460001

BT菌株への殺虫性タンパク質遺伝子導入ベクターの開発
科学研究費補助金(基盤研究(C))
1996 - 1996
浅野 真一郎
B.thuringiensisにおけるinsecticidal crystal protein(ICP)の発現制御機構が近年、いろいろなICP遺伝子について行われてきた。研究代表者は、B.thuringiensisのICP遺伝子発現ベクターを構築し、各種ICPを発現させることが可能となった。また、B.thuringiensis菌株から各ICP遺伝子のプロモーターをクローニングし(cry1A遺伝子プロモーター、cry2A遺伝子プロモーター、cry3A遺伝子プロモーター、cry4A遺伝子プロモーター等)各種プロモーター制御下でのそれぞれのICP遺伝子の発現制御機構について解析した。cry1Aとcry4A遺伝子に関しては、そのプロモーターでの発現制御機構が、詳細に調べられているがcry2A遺伝子に関してはこの遺伝子のプロモーター制御下でのICP遺伝子発現制御機構は調べられていなかった。cry2A遺伝子プロモーター発現制御機構を調査した結果、cry2A遺伝子の発現は、その上流域に存在する2つのorfの上流に存在するB.thuringiensisの他のcry遺伝子にも共通して見られる、BtIプロモーター制御下で発現されることがプライマーエクステンション法を用いた調査で明かとなった。crylA遺伝子プロモーターの発現制御機構と併せて、新たにcry2A遺伝子プロモーター発現制御機構について本実験で明らかにし、B.thuringiensis遺伝子導入ベクターを2種類構築することができた。
文部科学省, 基盤研究(C), 北海道大学, Competitive research funding, 08660063

Molecular mechanism on the replication of Bombyx densovirus
Grants-in-Aid for Scientific Research
1995 - 1996
BANDO Hisanori, SAHARA Ken, ASANO Shinichiro
Bombyx mori denso nucleosis virus (Yamanashi isolate ; BmDNV-2) is a small spherical virus with a bi-partite genome. In this study, to elucidate the nature of BmDNV-2, the viral DNA replication mechanism, genome organization and function of viral proteins were investigated. Analysis of the replicative intermediates (RI) of the viral DNA suggested the self-priming and hairpin-transfer model for parvovirus replication could not be applied to the replication of BmDNV-2. Nucleotide sequencing studies of the genomic DNAs showed that, unlike BmDNV-1 and the other parvoviruses the BmDNV-2 terminal sequences did not contained the large palindromic sequence which could form a stable hairpin structure. This was confirmed by the secondary structure prediction using a computer analyzes program. These results also suggested that the BmDNV-2 had a unique replication mechanisms other than the self-priming and hairpin-transfer mechanism.
Furthermore, the sequence analyzes elucidated that the BmDNV-2 genome contained 6 major open reading frames (ORFs), 4 ORFs in VD1 and 2in VD2. Immunochemical studies demonstrated that the major four viral structural proteins (VP1,2,3 and 4) were encoded in the ORF2 of VD1. Followingly, the functions of the nonstructural proteins (NSPs) were investigated by transient expression assay using the luciferase as a reporter. The NSPs, p37 and p14 which were encoded in ORF3 and4 of VD1, respectively, transactivated the tentative promoter sequence for the major structural protein gene, suggesting that these proteins were concerned in the regulation of the expression fo structural proteins. In addition, p27 which was an NSP encoded in VD2 could transactivate all of the viral promoters implying that p27 is a key regulatory protein in the BmDNV-2 replication.
In conclusion, this study elucidated the genome structure and the genome organization of the BmDNV-2. The results revealed that the BmDNV-2 is a new type of single-stranded DNA virus with a unique replication mechanism which was different from those of parvoviruses. Furthermore, the functional analyzes of the viral proteins suggested that the viral gene expression was regulated in the complicated manner by the proteins encoded in viral DNAs. And, p27 will be a key regulatory protein in the replication of the BmDNV-2 and may be one of the factor concerning in the determination of the host specificity.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (B), HOKKAIDO UNIVERSITY, 07456032

DNA probe synthesize for identification of cry genes from Bacillus thuringiensis which has a broad spectrum in the insecticidal crystal protein.
Grants-in-Aid for Scientific Research
1994 - 1996
IIZUKA Toshihiko, SAHARA Ken, ASANO Shinichiro, BANDO Hisanori
Becillus thuringiensis produces parasporal crystal protein which has insecticidal activity. This protein has been expressed by cry genes from B.thuringiensis. In the research. origonucleotide primers were synthesized depending on each cry gene specific region and DNA probes were prepared by PCR method. According to apply this method. cry genes from B.thuringiensis were easily and quickly isolated and identified.
We have also isolated B.thuringiensis strains from soil samples in Hokkaido and Indonesia. Cry genes from these isolates were identified by using PCR method and a couple of novel cry genes were analyzed for nucleotide sequencing. By the result. cryV gene which has both of Lepidopteracidal and Coleopteracidal activities were found from serovar kurstaki INA-02. These results should be applied for microbial control agents in the near future.
Japan Society for the Promotion of Science, Grant-in-Aid for Scientific Research (A), HOKKAIDO UNIVERSITY, 06404010

Isolation of BT and analysis of ICP gene in Indonesia.
Grants-in-Aid for Scientific Research
1995 - 1995
IIZUKA Toshihiko, HASTOWO Sugyo, LAY Bibiana W., SAHARA Ken, ASANO Shinichiro, BANDO Hisanori
Bacillus thuringiensis(Bt)is a gram-positive soil bacterium distinguished from other bacilli by its production of insecticidal protein. Because of their strong insecticidal activity and highly restricted host specificity, Bt has been used for microbiological control of plant-destroying insect. Recently, new Bts with unique characteristic traits have been isolated from various area in the world. Aim of our project is to isolate Bts with unique characteristics from Indonesia where is the habitant of huge number of insect species. In the last two years, we isolated many Bts from the soil collected from more than 50 areas in Indonesia. This year, we tried to obtain further Bts with unique properties from the areas where we have been able to obtain Bts before and characterized them.
We newly obtained 17 Bt strains from 6 areas including forests and sericultural farms, resulting in obtaining more than 80 Bt isolates in the three years-collaboration. In the H-serotyping, the isolates were classified into three subspecies, alesti, kenyae and indiana. It was demonstrated by the following researches such as analysis of ICP by SDS-PAGE,ICP gene-typing by PCR method established in our Lab, and bio-assay of the host specificity that our Bt collection contained three unique isolates which had never been reported. The results from the studies on the properties of these three unique Bt isolates lead us to the tentative conclusions that the minor mutation in the ICP gene results in the change of host specificity and that there is new ICP gene which has quite different nucleotide sequence in nature.
Japan Society for the Promotion of Science, Grant-in-Aid for international Scientific Research, Hokkaido University, 07044173

Isolation of BT and analysis of ICP gene in Indonesia.
Grants-in-Aid for Scientific Research
1993 - 1994
IIZUKA Toshihiko, LAY Bibiana w., SAHARA Ken, ASANO Shinichiro, BANDO Hisanori
Bacillus thuringiensis (BT) is known as the bacteria group which produces the insecticidal crystal proteins (ICP) and has been used for microbiological control of plant-destroying insect because of its highly restricted host specificity. Nowadays, new Bts with unique characteristic traits have been isolated from various areas in the world. Aim of our project is to isolate Bts from some areas in Indonesia where is a habitant of huge numbers of insect species. In the last year, we isolated many Bts from the soil under big trees as reported. This year, we tried to obtain further Bts from several areas and studied its biological characteristics.
As a result, we obtained more than 30 BT isolates form about 50 areas containing forest and sericultural field. In the H-serotyping, the isolates were classified into subspp. thuringiensis, sotto, darmstadiensis, aizawai but some were untypable. In the two years-collaboration, we obtained more than 60 BT isolates already and analyzed gene construction especially on crystal protein (cry) gene by PCR method established in our laboratory. Furthermore, the host specificity of the isolates were studied. The results of the experiments showed that the classification by H-serotype is not always reflected in the grouping of BT by its host spectrum. For example, some isolates obtained from Indonesia had the insecticidal activity against Spodoptera litura but it was classified into subsp. kurstaki which was a group of BT without insecticidal activity against the insect. One of those isolates, INA02, had the cryV gene which seemed to have an activity against S.litura. Then we cloned and determined the nucleotide sequence of the cryV gene of INA02.
On the other hand, we found another new type of BT which had the insecticidal activity against Aedes japonicus. We identified a cryII gene in this isolate (SKW isolate) which showed the sequence homology to the cryIIA gene. The function of this gene is under investigation. Another isolate (INA67) which was classified into subspp. tenebrionis showed the insecticidal activity against a coleoptera. However, this isolate contained no cryIII or V gene which were reported as the coleopteracidal crystal protein genes.
Japan Society for the Promotion of Science, Grant-in-Aid for international Scientific Research, Hokkaido University, 05044116

家蚕中腸上皮細胞上のdelta-エンドトキシン受容体に対するモノクローナル抗体作製
科学研究費補助金(奨励研究(A))
1993 - 1993
浅野 眞一郎, 浅野 真一郎
家蚕中腸上皮細胞上に存在する、Bacillus thuringiensis subsp.sottoのdelta-エンドトキシンにたいする受容体についてまず解析を進めた。まず家蚕中腸の組織切片を作成し、切片上にてdelta-エンドトキシンとのバインディングアッセイを行ったところ、家蚕中腸上皮細胞にdelta-エンドトキシンに強い親和性を持つ受容体が存在していることが明らかとなった。Wolfersbergerらの方法に従い、Mg/EDTA沈殿と分画遠心法によって調整した家蚕中腸からBrush border membrane vesicle(BBMV)を調整し、SDS-P AGEによる分画の後、ウエスタンブロットしsotto由来のdelta-エンドトキシンとのバインディングアッセイによる解析によって、BBMV由来の幾つかの分画のバンドとバインディングが認められた。その中でも役130kdalのバンドに強いバインディングが認められたので、そのバンドの分画をSDS-ポリカクリルアミドゲルから切り出して電気泳動的にその分画のペプチドを回収した。その回収した分画のペプチドとdelta-エンドトキシンとをco-incubateし十分にそのペプチドとdelta-エンドトキシンとバインディングさせた後、ウェスタンブロットしたBBMV分画とのバインディングアッセイを行ったところ、バインディングが押さえられていることが明らかとなった。よって、家蚕中腸BBMVをSDS-PAGEによって分画して、ゲルから回収し、その回収した分画をマウスに皮下接種して現在免疫化している。それによって得られた抗体を用いて、家蚕BBMVとsotto由来のdelta-エンドトキシンとのバインディングを阻害することができるかを確かめるつもりである。
文部科学省, 奨励研究(A), 北海道大学, Principal investigator, Competitive research funding, 05856008

家蚕濃核病ウイルスの遺伝子構造と増殖機構
科学研究費助成事業
1992 - 1992
伴戸 久徳, 浅野 真一郎
家蚕濃核病ウイルスには現在、伊那株(BmDNV-1)と山梨株(BmDNV-2)に代表される2種類のウイルスが存在し、これらは品種特異性などの点で著しく異なっている。本研究ではこれらのウイルスの違いを分子生物学的な基盤に於いて明らかにするとともに、濃核病発生予防のための基礎的情報を得る目的で以下の実験を行った。
BmDNV-1はすでにその遺伝子構造及び複製様式からパルボウイルスに属する事が判明している。ところが、BmDNV-2に於いてはゲノム末端にヘアピン構造は認められず複製様式に於いてパルボウイルスとは異なっている事が推定された。本ウイルスの分類上の位置づけを明瞭にするためにはウイルスDNAの遺伝情報に関して明らかにする必要がある。そこで2種類のゲノムDNA(VD1,VD2)の塩基配列の解析を行った。まず、VD2に関してその全塩基配列を決定した。VD2は約6000塩基からなりその塩基組成には片寄りがみられた。すなわち末端領域は非常にG-Crichでありそれ以外の領域は逆に非常にA-Trichであった。また、本配列には少なくとも2個の大きなオープン・リーディング・フレーム(ORF)が認められ、これらは別々のDNA鎖に存在した。これらのORFにコードされるタンパク質に関して解析を進めるためそれらに対するペプチド抗体を作成中である。VD1に於いてもVD2同様塩基配列の片寄りが認められた。現在、遺伝情報の解析中である。以上の結果は、BmDNV-2がこれまで知られているDNVとは全く異なるタイプのウイルスである事を示している。一方、BmDNV-1とBmDNV-2に於いて明かとなった塩基配列を基に合成したオリゴヌクレオチドを用いたPCR法の応用を検討したところ、極めてて高感度でそれぞれの塩基配列が検出可能であり、この方法の診断及び疫学への応用が可能であると考えられた。
日本学術振興会, 一般研究(C), 北海道大学, 04660057

Analysis of nucleotide sequence of insecticidal crystal protein (ICP) gene which shows highly activity against Lepidoptera insect.
Grants-in-Aid for Scientific Research
1991 - 1992
IIZUKA Toshihiko, KIKUTA Harunori, ASANO Shinichiro, BANDO Hisanori
A insecticidal crystal protein (ICP) produced by Bacillus thuringiensis (BT), which shows highly activity against the Lepidopteran insect, should be studied from the following two ways:
1. Newly isolation and identification of BT. : Newly BT isolates from soil samples which were collected in the natural conserved area, were identified by routine technique of H-serotype and by newly developed technique depending on agarose gel electrophoresis of cryl genes. After oligo-primers designed by a specific domain were synthesized, these cryI genes were amplified by PCR method with template DNA from BT subspecies. The newly identification technique was very effective in order to clear a high active strain comparing with already known BT strains.
2. Nucleotide sequence analysis of ICP genes. : BT subsp. wuhanensis has been known as high active strain against the silkworm and the cabbage moth. According to a report of Iizuka et al. (1993), subsp. wuhanensis has included cryIA(b), cryIA(c) and cryID genes. In this study, the nucleotide sequence of cryIA(b) was analyzed and compared with another cryIA(b) genes from subspp. thuringiensis berliner 1715 and kurstaki HD-2. The differentiation of amino acid composition between wuhanensis and HD-2 was only one within a active domain. A regulatory mechanisms of each protein expression encoded by three genes was in progressed.
Japan Society for the Promotion of Science, Grant-in-Aid for General Scientific Research (B), Hokkaido University, 03454058

殺虫活性を有する蛋白質、その蛋白質をコードするDNA、有害生物防除剤及び防除方法。
Patent right, 浅野 眞一郎, 山中 聡, 武内 克義, 株式会社エス・ディー・エス バイオテック
特願2000-236140, 03 Aug. 2000
特開2002-045186, 12 Feb. 2002
特許第4608059号
15 Oct. 2010
200903052090260338

ドウガネブイブイの防除方法及び防除剤
Patent right, 浅野 眞一郎, 松木 伸浩, 西橋 秀治, 木村 雅敏, 江原 岳, DIC株式会社
特願平11-338627, 29 Nov. 1999
特開2001-151617, 05 Jun. 2001
特許第4337193号
10 Jul. 2009
200903010080881461