马尾松毛虫CPV基因组第7片段的cDNA克隆及序列分析  

Molecular Cloning and Sequence Analysis of Dendrolimus punctatus Cytoplasmic Polyhedrosis Virus (DpCPV) Segment 7

作　　者：汪洋[1] 张珈敏[1] 李杨[1] 李艳秋[1] 胡远扬[1] 

机构地区：[1]武汉大学病毒研究所,湖北武汉430072

出　　处：《武汉大学学报（理学版）》2004年第2期216-222,共7页Journal of Wuhan University:Natural Science Edition

摘　　要：利用琼脂糖凝胶电泳分离马尾松毛虫质型多角体病毒(DpCPV)基因组dsRNA,并回收纯化第7片段(S7).经逆转录和PCR扩增,获得2个亚克隆片段并测序.再根据已测序列设计引物,利用RNA连接酶介导的RACE法得到S7两端克隆并测序.通过拼接可得到DpCPVS7全序列.经过序列分析发现,S7全长1502bp,5′端具有CPV 1型末端保守序列AGTAA,3′端具有保守序列GTTAGCC.起始密码子ATG位于25～27残基,终止密码子TAG位于1373～1375残基,推测S7片段编码448个氨基酸多肽,分子量约为5.0×107.与舞毒蛾质型多角体病毒(LdCPV 1)和家蚕质型多角体病毒(BmCPV)第7片段相比较,核苷酸同源性分别为99%和86%.Segment 7 dsRNA of DpCPV was purified by extraction kit after being excised from the gel. Two sub-clone segments were obtained by RT-PCR using the primers constructed on the basis of homologous sequences. Primers were constructed on the basis of the 2 sub-clone segments' sequences and the sequences of both terminal parts of S7 were obtained by RLM-RACE. Following splicing, the full-length sequence of DpCPV Segment 7 was obtained. Analysis on the sequence of S7 revealed that it consists of 1 502 nucleotides encoding putative protein of 448 amino acids with molecular mass of 5.0×10~7. Comparison of the nucleotide sequence of the segment 7 of DpCPV with that of BmCPV and LdCPV-1 showed that nucleotides homology is 86%and 99%, respectively.

关 键 词：马尾松毛虫 质型多角体病毒 CDNA末端快速扩增 CPV基因组 CDNA克隆 序列分析 呼肠孤病毒科 

分 类 号：S432.41[农业科学—植物病理学] Q78[农业科学—农业昆虫与害虫防治]