Molecular Basis for Safety Assessment of Microbial Pesticides

微生物农药安全评估的分子基础

• 批准号: 15208006
• 负责人: BANDO Hisanori
• 金额: $ 32.61万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15208006/
• 关键词: Baculovirus; Bacillus thuringiensis; Host specificity; Enterotoxin; Entomopathogenic microorganisms; マイクロアレイ; プロモーター; バチルスチューリンゲンシス; コアプロモーター; バチルスセレウス; ie1

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.

杆状病毒和Bt（苏云金芽孢杆菌）在世界各地被用作微生物农药，前者还提供了真核生物中最有效的外源基因表达系统（杆状载体系统）。我们研究了这些昆虫病原微生物安全性的分子基础。最近的研究表明，杆状病毒（AcMNPV）可以独立于细胞周期进入哺乳动物细胞核而不进行增殖，并且可以有效表达外源基因而不表达自身基因，这表明杆状病毒可以作为基因治疗领域的低风险病毒载体。与一般看法相反，我们阐明了 AcMNPV 至少部分通过通常的感染途径在哺乳动物细胞中表达病毒基因。此外，接种AcMNPV的哺乳动物细胞中细胞基因（至少β-肌动蛋白）的表达上调。然而，在哺乳动物细胞中，病毒基因的表达部分通过组蛋白去乙酰化机制被抑制到较低水平，并且进一步导致一些病毒基因的转录在错误的位点开始。对于苏云金芽孢杆菌（Bt）菌株，我们主要研究了非溶血性肠毒素C（NheC）对Vero细胞的毒性机制，该毒素被认为是最重要的肠毒素引起的。 食物中毒。研究表明，具有Vero细胞毒性的蜡状芽孢杆菌（Bc）菌株临床分离株分泌的NheC可以通过N端30个氨基酸区域附着在Vero细胞表面。另一方面，商业产品中使用的 Bt 菌株分泌的 NheC 中，相应的 N 末端区域可能被翻译后切割所截短。此外，NheC的N末端截短不再表现出Vero细胞毒性。

项目成果

The W chromosome detection in several lepidopteran species by genomic in situ hybridization (GISH)

基因组原位杂交 (GISH) 检测几种鳞翅目物种的 W 染色体

• 发表时间: 2006
• 期刊: J Insect Biotech Seric 75
• 作者: Yoshido A;Yamada Y;Sahara K
• 通讯作者: Sahara K

Bacillus thuringiensis distribution in soil of Vietnam

越南土壤中苏云金芽孢杆菌的分布

• 发表时间: 2005
• 期刊: Biotechnology of Bacillus thuringiensis 5
• 作者: Miyadai;H.;N. D. Binh
• 通讯作者: N. D. Binh

Marked decrease of ribosomal RNA in BmN cells infected with AcMNPV

感染 AcMNPV 的 BmN 细胞中核糖体 RNA 显着减少

• 发表时间: 2005
• 期刊: J. Insect Biotech. Sericol. 74・3
• 作者: Yoshinaga;K.;Arimura;SI.;Niwa;Y.;Tsutsumi;N.;Uchimiya;H.;Kawai-Yamada;M.;R. Fujita
• 通讯作者: R. Fujita

DNA microarrays of baculovirus genomes: differential expression of viral genes in two susceptible insect cell lines

• DOI: 10.1007/s00705-002-0922-3
• 发表时间: 2003-03
• 期刊: Archives of Virology
• 影响因子: 2.7
• 作者: Junya Yamagishi;R. Isobe;T. Takebuchi;H. Bando

The Bombyx mori karyotype and the assignmet of linkage groups

家蚕核型及连锁群分配

• 发表时间: 2005
• 期刊: Genetics 170・2
• 作者: Kovacs;J.J.;Murphy;P.J.M.;Gaillard;S.;Zhao;X.;Wu;J.-T.;Nicchitta;C.V.;Yoshida;M.;Toft;D.O.;Pratt;W.B.;Yao;T.-P.;A. Yoshido
• 通讯作者: A. Yoshido