运用Codon W、ClustalX、TreeView软件及EMBOSS(The European Molecular Biology Open SoftwareSuite)、CIMMiner在线分析软件对选取的29株PRRSV ORF1a基因进行密码子偏爱性聚类分析.CAI、CBI、Fop、Nc、GC3s和GC含量、基因长度等相关...运用Codon W、ClustalX、TreeView软件及EMBOSS(The European Molecular Biology Open SoftwareSuite)、CIMMiner在线分析软件对选取的29株PRRSV ORF1a基因进行密码子偏爱性聚类分析.CAI、CBI、Fop、Nc、GC3s和GC含量、基因长度等相关性分析显示PRRSV各毒株编码的ORF1a基因密码子偏爱性各有差异,其中Lelystad virus、LV4.2.1、VR-2332、RespPRRS MLV与国内分离的高致病性PRRSV变异株之间差异较大.密码子使用概率聚类分析表明CC-1、NVSL-97-7895、CH-1a、RespPRRS MLV、LV4.2.1、Lelystad virus与高致病性PRRSV变异株距离较远,而国内分离株相互间的聚类距离则较接近,此结果与基于氨基酸序列比对构建的系统进化树图谱基本一致.由此可见,PRRSV病毒ORF1a基因密码子使用偏爱性的差别与病毒的遗传多样性密切相关.展开更多
A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuc...A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuclectide sequence was determined by the dideoxy mediated chain termination method.The results showed that the complete open reading frame (ORF) of ORF1 gene encoding 314 amino acids was 945 bp in length.A comparison of the nucleotide and amino acid sequences of ORF1 gene with that of other PCV strains showed that the identity of nucleotide with PCV 1 and PCV 2 were 83% and 96 4%～99 2% respectively,and identity of the deduced amino acid with PCV 1 and PCV 2 were 84% and more than 98% respecitively.The DNA fragment of ORF1 gene was subcloned into prokaryotic expression vector pET 28a and pGEX KG;while the specific non fusion and fusion proteins with GST of molecular weight 38 kD and 63 kD were expressed in E.coli BL 21 (DE3).Western blotting assay indicated that the polyclonal antibody against PCV 2 could recognize these two proteins.展开更多
文摘A pair of specific primers were designed and synthesized according to the published sequences of ORF1 gene of PCV 2. The complete DNA fragment of ORF1 gene was obtained by PCR from viral DNA of PCV 2 QD strain.Its nuclectide sequence was determined by the dideoxy mediated chain termination method.The results showed that the complete open reading frame (ORF) of ORF1 gene encoding 314 amino acids was 945 bp in length.A comparison of the nucleotide and amino acid sequences of ORF1 gene with that of other PCV strains showed that the identity of nucleotide with PCV 1 and PCV 2 were 83% and 96 4%～99 2% respectively,and identity of the deduced amino acid with PCV 1 and PCV 2 were 84% and more than 98% respecitively.The DNA fragment of ORF1 gene was subcloned into prokaryotic expression vector pET 28a and pGEX KG;while the specific non fusion and fusion proteins with GST of molecular weight 38 kD and 63 kD were expressed in E.coli BL 21 (DE3).Western blotting assay indicated that the polyclonal antibody against PCV 2 could recognize these two proteins.
