Production of Transgenic Silkworms Resistant to BmNPV

抗 BmNPV 转基因蚕的生产

• 批准号: 12306002
• 负责人: BANDO Hisanori
• 金额: $ 29.89万
• 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: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12306002/
• 关键词: silkworm; RNAi; NPV; virus resistance; piggy Bac; DNA Microarray; primase; helicase; ウィルス耐性; トランスジェニック; BmNPV; IE1; アンタゴニスト; トランスポゾン

The nuclear polyhedorosis disease caused by BmNPV infection is a major problem in suriculture around the world. The inhibition of the functional expression of these essential genes in cells infected with NPV should be an effective strategy for preventing mortal infections of BmNPV in the silkworm.An immediate-early gene product of baculovirus, IE1, is essential for viral gene expression and for viral DNA replication. We constructed an antagonistic IE1 (IE1TN) of Bombyx mori nuclear polyhedrosis virus (BmNPV)s and demonstrated markedly reduced virus production in BmN cells expressing IE1TN. On the other hand, recent study demonstrated that dsRNA is a powerful tool for gene-specific silencing in plants and animals. Then, we constructed recombinant plasmids designed for expressing dsRNAs with the sequences of the essential viral gene, such as Ief-1 and dnahel and demonstrated that viral production was effectively inhibited in BmN cells expressing dsRNAs. Finally, using a transposon piggyback system, we confirmed the moderate BmNPV-resistance caused by the transgenesis of the IE1TN-or dsRNA-expressing gene in the transgenic silkworms.

由BmNPV感染引起的核性多角症是世界范围内水产养殖的一个主要问题。抑制NPV感染细胞中这些必需基因的功能表达可能是预防家蚕BmNPV致命感染的有效策略。杆状病毒的一种即时早期基因产物IE1对病毒基因表达和病毒DNA复制至关重要。我们构建了家蚕核型多角体病毒（Bombyx mori nuclear polyhedrosis virus, BmNPV） 5的拮抗IE1 (IE1TN)，并证实在表达IE1TN的BmN细胞中显著降低了病毒的产量。另一方面，最近的研究表明，dsRNA是植物和动物基因特异性沉默的有力工具。然后，我们构建了含有必要病毒基因Ief-1和dnahel序列的表达dsRNAs的重组质粒，并证明了在表达dsRNAs的BmN细胞中，病毒的产生被有效抑制。最后，利用转座子背带系统，我们证实了ie1tn或dsrna表达基因在转基因家蚕中引起的中度bmnpv抗性。

项目成果

S.Matsumoto: "Characterization of acyl-CoA-Binding protein (ACBP) in the pheromone gland of the silkworm, Bombyx mori"Insect Biochemistry and Molecular Biology. 31. 603-609 (2001)

S.Matsumoto：“家蚕信息素腺中酰基辅酶 A 结合蛋白 (ACBP) 的表征”昆虫生物化学和分子生物学。

K.Sahara: "Study on the novel forage tree for Antheraea yamamai Guerin"Applied Entomology and Zoology. 45. 94-98 (2001)

K.Sahara：“Antheraea yamamai Guerin 新型饲料树的研究”应用昆虫学和动物学。

<Tamura, T.>_-, Quan, G.X., Kanda, T., Kuwabara, N.: "Transgenic silkworm research in Japan : Recent progress and future"Proceeding of Joint International Symposium of Insect COE Research Program and Insect Factory Research Project. 77-82 (2001)

<Tamura, T.>_-, Quan, G.X., Kanda, T., Kuwabara, N.：“日本转基因蚕研究：最新进展和未来”昆虫 COE 研究计划和昆虫工厂研究项目联合国际研讨会论文集

K.Sahara: "W-derived BAC probes as a new tool for identification of the W chromosome and its aberrations in Bombyx mori"Chromosoma. 112. 48-55 (2003)

K.Sahara：“W 衍生的 BAC 探针作为鉴定家蚕 W 染色体及其畸变的新工具”。

GX.Quan: "Characterization of the kynurenine 3-monooxygenase gene corresponding to the white egg 1 mutant in the silkworm Bombyx mori"Molecular Genetics and Genomics. 267. 1-9 (2002)

GX.Quan：“家蚕 Bombyx mori 中白卵 1 突变体对应的犬尿氨酸 3-单加氧酶基因的表征”分子遗传学和基因组学。