茶蚕颗粒体病毒AcMNPVORF38同源基因的克隆及序列分析

目的]为利用昆虫病毒更好地防治害虫和进行分子生物学研究。方法]从感病茶蚕幼虫的尸体中提取颗粒体病毒DNA后,进行酶切、克隆、测序和序列分析。结果]成功克隆了茶蚕颗粒体病毒基因组中一个1 484 bp的片段。该片段含有1个完整的开放阅读框(ORF)和2个不完整ORF。完整ORF为666 bp,有典型Nudix结构域,其编码1个与苜蓿银纹夜蛾核多角体病毒AcMNPV的ORF38高度同源的基因,定名为AcORF38同源基因。2个不完整ORF分别编码p47蛋白基因的C-端和p24基因的N-端。结论]茶蚕颗粒体病毒与卷蛾科宿主中颗粒体病毒亲源关系较近,而与夜蛾科宿主昆虫中颗粒体病毒较远。AcORF38同源基因与编码NTP焦磷酸水解酶的基因高度同源,可能与病毒的复制和DNA损伤的修复过程相关,也可能与入侵寄主的过程相关。

Cloning and Sequence Analysis of AcMNFVORF38 Homologous Gene in Granulovius of Andraca bipunctata Walker

Objective] The research aimed to better understand the molecular biology of insect viruses.Method] After virus DNA was extracted from the cadaver of diseased larva of Andraca bipunctata,it was made for enzyme digestion,cloning,sequencing and sequence analysis.Result] A fragment with the size of 1 484 bp in the genome of granulosis virus in A.bipunctata was successfully cloned.This fragment contained one complete open reading frame (ORF) and 2 incomplete ORF.The complete ORF was 666 bp and had the typical Nudix domain,coding one gene which was highly homologous with ORF38 of Autographa californica nucleopolybedrovirus AcMNPV and named as AcORF38 homologous gene.2 incomplete ORF coded C terminal of p47 gene and N-terminal of p24 gene.The granulosis virus in A.bipunctata had closer genetic relationship with that in Tortricidae host,while it had farther genetic relationship with host insects of Noctuidae.Conclusion] AcMNPV-ORF38 homologous gene was highly homologous with the gene coding NTP pyrophosphate hydrolase,which was probably related to viral replication and the repairing process of DNA damage,or related with the process of invading hosts.