猪水泡病病毒5''和3''非编码区一级和二级结构分析

应用RT-PCR技术,用特异性引物扩增出SVDV HK’70 5’和3’非编码区,并将目的片段连接于pMD18-T载体,转化JM109感受态细胞。重组质粒经双酶切、PCR鉴定后测序。核苷酸序列比较结果表明,SVDV HK’70 5’非编码区与参考毒株SVDV J1’73、SVDV H/3’76核苷酸的差异率分别为1.6％、1.9％;SVDV HK’70 3-NCR与参考毒株SVDV J1’73、SVDV H/3’76核苷酸的差异率分别为1.0％、1.0％。应用计算机软件对3个SVDV毒株非编码区的二级结构进行预测,结果表明,SVDV HK’70 5’非编码IX-"级结构中存在13个结构域,SVDV J1’73 5’非编码区二级结构中存在9个结构域,SVDV H/3’76 5’非编码区二级结构中存在10个结构域;SVDV HK’70 3’非编码区二级结构中存在3个结构域,且与其他的两个毒株SVDV J1’73、SVDV H/3’76基本相同。

The primary and secondary structure analysis of 5''''and 3'''' noncoding regions of swine vesicular disease virus

Amplicating SVDV HK' 70 5' and 3' nonceding region by RT-PCR, DNA fragment ligating with pMD18-T and transformed JM109. Recombinant plasmid was identified with restriction enzyme and PCR. Sequence analysis indicated: SVDV HK'70 5' noncoding region DNA sequence diversity rates was 1.6 % with SVDV J1 '73,1.9 % with SVDV H/3' 76; SVDV HK'70 3' noncoding region DNA sequence diversity rates was 1.0 % with SVDV JI'73,1.0 % with SVDV H/3' 76.5' noncoding region secondary structure indicated: SVDV HK'70 secondary structure had thirteen structure regians, SVDV Jl '73 had nine secondary structure, SVDV H/3' 76 had ten secondary sturcture; 3' noncoding region secondary structure indicated: SVDV HK'70 secondary structure had three structure regions, and SVDV Jl'73, H/3' 76 had same secondary structure.