马尾松毛虫CPV基因组第7片段的cDNA克隆及序列分析

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 汪洋 WANG Yang 1 2 first-author 张珈敏 ZHANG Jia-min 1 2 李杨 LI Yang 1 2 李艳秋 LI Yan-qiu 1 2 胡远扬 HU Yuan-yang 1 2 武汉大学病毒研究所 武汉大学病毒研究所 Institute of Virology, Wuhan University Institute of Virology, Wuhan University 武汉大学病毒研究所,湖北,武汉,430072 武汉大学病毒研究所,湖北,武汉,430072 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. 马尾松毛虫 质型多角体病毒 cDNA末端快速扩增 Dendrolomus punctatus cytoplasimc polghedrosis virus rapid amplication of cDNA ends（RACE） Q78 2004-02-01 2021-04-01 2