斜纹夜蛾核型多角体病毒Ⅱ型DNA复制原点的功能分析  

Function of DNA Replication Origins from Spodoptera litura Nucleopolyhedrovirus Ⅱ

作　　者：刘惠芬[1] 刘文光[1] 衣葵花[1] 张志芳[2] 李云芝[1] 郭光[1] Liu Huifen;Liu Wenguang;Yi Kuihua;Zhang Zhifang;Li Yunzhi;Guo Guang(Sericultural Research Institute of Shandong,Yantai Shandong 264002,China;Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China)

机构地区：[1]山东省蚕业研究所,山东烟台264002 [2]中国农业科学院生物技术研究所,北京100081

出　　处：《蚕业科学》2019年第5期686-690,共5页ACTA SERICOLOGICA SINICA

基　　金：山东省自然科学基金项目(No.ZR2017BC080);山东省现代农业产业技术体系蚕桑产业创新团队建设项目(No.SDAIT-18-03,SDAIT-18-01)

摘　　要：斜纹夜蛾核型多角体病毒Ⅱ型(SpltNPVⅡ)基因组DNA含有7个同源重复区(hr)。采用脂质体介导转染法和实时荧光定量PCR对7个hr在异源病毒感染的BmN和Sf21细胞中的复制原点功能进行分析。结果表明,SpltNPVⅡhr在BmN和Sf21细胞中均具有复制原点功能,但其复制起始能力不同。BmNPV感染的BmN细胞中,hr4复制起始能力最强,每微克DNA中的拷贝数达405×10^5个,hr5复制起始能力最弱,仅为221×10^4个拷贝/μg;AcMNPV感染的Sf21细胞中hr1复制起始能力最强,可达592×10^6个拷贝/μg,hr5复制起始能力最弱,仅为191×10^4个拷贝/μg。另外,SpltNPVⅡhr在异源细胞中的复制起始能力与其所含回文序列及复制相关的基序等特征结构的数目存在显著的正相关。There were 7 homologous regions(hrs),namely Sphr1 to Sphr7 in the genomic DNA of Spodoptera litura nucleopolyhedrovirusⅡ(SpltNPVⅡ). Liposome-mediated transfection and real-time PCR were carried out to assess the function of these hrs as DNA replication origins(oris)in BmN and Sf21 cells infected with heterologous viruses. The results revealed that all hrs could function as ori of DNA replication in BmN and Sf21 cells,but had different replication ability,which varied from 4 05×10^5 copies(hr4)to 2 21×10^4 copies(hr5)in 1 μg DNA of BmN infected with BmNPV,and from 5 92×10^6 copies(hr1)to 1 91×10^4 copies(hr5)in 1 μg DNA of Sf21 infected with AcMNPV. In addition,replication ability of Sphr1-7 as ori in heterogeneous cells had significantly positive correlations with the number of characteristic sequences like palindromes and replication related motifs.

关 键 词：复制原点 同源重复区 斜纹夜蛾核型多角体病毒 

分 类 号：S884.51[农业科学—特种经济动物饲养] Q789[农业科学—畜牧兽医]