カイコ核多角体病ウイルス遺伝子の構造と機能の解析

家蚕核型多角体病毒基因结构与功能分析

• 批准号: 05660388
• 负责人: KOBAYASHI Michihiro
• 金额: $ 1.34万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05660388/
• 关键词: Silkworm; Nuclear polyhedrosis Virus; DNA polymerase; Gene; Expression

A gene encoding a putative DNA polymerase (pol) of Bombyx mori nuclear polyhedrosis virus (BmSNPV) was cloned and sequenced. The gene included an open reading frame (ORF) encoding a polypeptide of 988 amino acids with a predicted molecular mass of 114.65 kDa. The deduced amino acid sequence of the BmSNPV pol ORF showed an overall identity of 96 and 45% to those of the Autographa californica NPV (AcMNPV) pol ORF and the Lymantria dispar NPV pol ORF,respectively, and contained sequences conserved in a variety of eukaryotic and viral replicative DNA polymerases. The BmSNPV pol lacked a canonical TATAA element but contained a G+C-rich sequence in the transcriptional initiation region. Analyzes by Northern blot hybridization, RNase protection assay, primer extension, and 3' and 5' RACE (rapid amplification of cDNA ends) showed that at least seven different transcripts of approximately 3.1 kb that shared a common 3' end were expressed from the BmSNPV pol. The expression of these transcripts … More from BmSNPV pol was regulated differentially during virus infection. Transcription of five of the seven species initiated in the close vicinity of and within the motif 5'-GCGTGCT-3'. One transcript placed its initiation site within the motif 5'-AGAGCGT-3' and the remaining one within the motif 5'-GGCGGTGG-3'. The motifs 5'-GCGTGCT-3' and 5'-AGAGCGT-3' have been identified in pol and other genes of AcMNPV as conserved sequences containing transcriptional initiation sites, whereas the motif 5'-GGCGGTGG-3', which is arranged as a direct repeat in BmSNPV pol but not in AcMNPV pol, has not been defined as the sequence responsible for transcriptional initiation sites. The BmSNPV pol transcripts were detectable at 2 hr postinfection (p.i.) , peaked at 10 hr p.i., and declined to a low level by 18 hr p.i. The expression of BmSNPV pol was not inhibited but delayd dramatically by the protein synthesis inhibitor cycloheximide upon treatment of infected cells, whereas aphidicolin, an inhibitor of DNA polymerase, inhibited BmSNPV pol transcription. These results suggest a complicated and unique mechanism for the regulation of BmSNPV pol expression. Less

编码Bombyx Mori核多角质病毒（BMSNPV）的假定DNA聚合酶（POL）的基因被克隆并进行了测序。该基因包括编码988个氨基酸的多肽的开放式阅读框（ORF），预测分子质量为114.65 kDa。 BMSNPV POL ORF的推导氨基酸序列的总体身份分别为96％和45％的加利福尼亚NPV（ACMNPV）POL ORF和lymantria dispar npv polf Orf的氨基酸序列，并包含序列，并包含在各种真生性和病毒性dna Polymersases中构成的序列。 BMSNPV POL缺少规范的TATAA元素，但在转录起始区域中包含富含G+C的序列。通过Northern印迹杂交，RNase保护测定，底漆扩展以及3'和5'种族（cDNA末端的快速扩增）的分析表明，从BMSNPV POL表达了至少七个共有3'端的约3.1 KB的转录本。在病毒感染期间，这些转录本的表达……更多来自BMSNPV POL的调节。在基序5'-GCGTGCT-3'的近距离附近启动的七个物种中的五个转录。一个成绩单将其主动站点放置在图案5'-agagcgt-3'中，其余部分将其放置在图案5'-ggcggtgg-3'中。在POL和其他ACMNPV的其他基因中确定了5'-GCGTGCT-3'和5'-agagcgt-3'的主题，作为包含转录启动位点的配置序列，而序列5'- gggcggtgg-3'，而在bmsnpv pol中均未在acmnpv pol中直接重复，但在acmnpv pol中均未授权。倡议网站。在感染后2小时（P.I.）检测到它们，在10小时的峰值峰值，并在18小时下降到低水平。 BMSNPV POL的表达不会被抑制，而是通过治疗感染细胞的蛋白质合成抑制剂环己酰亚胺的延迟延迟，而DNA聚合酶的抑制剂蚜虫抑制了BMSNPV POL转录。这些结果暗示了调节BMSNPV POL表达的复杂而独特的机制。较少的

项目成果

Sotoshiro, H.and Kobayashi, M.: "Identification of viral structural polypeptides in the midgut and feces of the silkworm, Bombyx mori, infected with Bombyx densovirus type 2." J.Invertebr.Pathol.(in press).

Sotoshiro, H. 和 Kobayashi, M.：“感染 2 型家蚕浓核病毒的家蚕中肠和粪便中病毒结构多肽的鉴定。”

Chaeychomsri, S., Ikeda, M.and Kobayashi, M.: "Nucleotide sequence and transcriptional analysis of the DNA polymerase gene of Bombyx mori nuclear polyhedrosis virus." Virology. 206. 435-447 (1995)

Chaeychomsri, S.、Ikeda, M. 和 Kobayashi, M.：“家蚕核多角体病毒 DNA 聚合酶基因的核苷酸序列和转录分析。”

Chaeychomsri, S., Ikeda, M., Kobayashi, M.: "Expression of viral DNA polymerase gene in the larvae of the silkworm, Bombyx mori (Lepidoptera : Bombycidae), during the infection with nuclear polyhedrosis virus." Appl.Entomol.Zool.(in press).

Chaeychomsri, S.、Ikeda, M.、Kobayashi, M.：“在感染核型多角体病毒期间，家蚕（鳞翅目：家蚕科）幼虫中病毒 DNA 聚合酶基因的表达。”

Kobayashi,M.and Caeychomsri,S.: "Thermal inhibition of viral disease in the silkworm,Bombyx mori" Indian Journal of Sericulture. 32. 1-7 (1993)

Kobayashi,M. 和 Caeychomsri,S.：“家蚕病毒性疾病的热抑制，Bombyx mori”《印度蚕业杂志》。

Kobayashi, M.and Chaeychomsri, S.: "Thermal inhibition of viral disease in the silkworm" Bombyx mori.Indian J.Sericult.32. 1-7 (1993)

Kobayashi, M. 和 Chaeychomsri, S.：“蚕病毒性疾病的热抑制”Bombyx mori.Indian J.Sericult.32。