摘要
以本实验室分离鉴定犬细小病毒新疆石河子株(CPV-SHZ)的DNA为模板,根据基因库已发表CPV序列设计合成了VP2基因的1对特异引物,进行聚合酶链式反应,扩增出约1.7kb的片段,按常规方法克隆进pMD18-T载体,经EcoR I和Sal I双酶切筛选到阳性质粒。测序得到VP2全基因组序列,并登陆Genbank(EU170352)。进一步对该片段进行序列分析,结果表明:所扩增基因片段长度为1755bp,与CPV参考株毒株V154(Type2a)、LCPV-V204(Type2b)、LCPV-V139(Type 2c(a))、LCPV-V203(Type 2c(a)),其核苷酸的同源性分别为99.32%、98.75%、98.97%、98.69%,确定CPV-SHZ株基因型为2a型,将其同我国和世界其他国家主要分离株进行基因系统发生进化关系分析,结果表明,其与我国北京分离株BJ018/07亲缘关系较近。
A pair of primers was designed and synthesized based on VP2 gene of Canine parvovirus （CPV） from GenBank. The genomic DNA samples of CPV strain isolated in Xinjiang were extracted and used as templates for polymerase chain reaction to amplify the VP2 gene. The specific 1755bp band was amplified and cloned into pMD18-T vector. The recombinant plasmids were identified by restriction endonuclease analysis. The exogenous insert of recombinant plasmid was further confirmed by sequence analysis. After sequencing, submission of sequence data to GenBank and the accession number is（EU170352）. The VP2 gene of Xinjiang isolation was compared with that of reference strains V154（Type2a）, LCPV-V204（Type2b）, LCPV-V139（Type 2c （a） ）, LCPV-V203 （Type 2c （a） ）. The nucleotide homology respectively is 99.32% ,98.75% ,98.97% ,98.69% and classified as a CPV-2a type. Compared with strains isolated in China and other parts of the world, the homology closer with strain BJ018/07.
出处
《石河子大学学报（自然科学版）》
CAS
2008年第3期311-315,共5页
Journal of Shihezi University(Natural Science)
