猪瘟病毒石门株动物传代脾毒与组织培养细胞毒E2基因序列的分析

Sequencing analysis of E2 gene of muti-passsaging spleen virus and cytopathogenic clsssical swine fever virus

根据已发表的猪瘟病毒基因组序列,设计合成了2对引物,以脾毒和细胞毒总RNA为模板,利用反转录聚合酶链式反应(RT-PCR)、套式聚合酶链式反应(nPCR)及测序技术,对多次动物传代和致细胞病变的猪瘟病毒主要囊膜糖蛋白E2基因的核苷酸序列进行了测定,用DNAstar软件比较分析了E2基因的核苷酸和氨基酸序列的同源性。结果发现,脾毒和标准石门株核苷酸序列的同源性为99.4%,氨基酸序列的同源性为99.0%;细胞毒与标准石门株核苷酸序列的同源性为98.3%,氨基酸序列的同源性为96.7%。由此说明,猪瘟病毒经猪体多次传代和组织培养致细胞病变后,均能保持遗传的稳定性。

<Abstrcat>According to the published CSFV sequence,two pairs of primers were designed.E2 gene of muti-passsaging and cytopathogenic clsssical swine fever virus was amplified and sequenced through RT-PCR,nPCR method.The nucletide sequences and the deduced amino sequences of E2 gene were compared and analyzed by DNA star software.The results showed that the nucleotide homology between spleen virus and Shimen was 99.4%,the aminoacid homology 99.0%,the nucleotide homology between cultured cell virus and Shimen 98.3%,the aminoacid homology 96.7%.So it indicates that E2 genes of CSFV can be genetically stable after muti-passsaging and cytopathogenic effect.