猪瘟病毒石门株在ST细胞连续传代后E0基因变异分析

为了解猪瘟病毒(CSFV)石门(Shimen)株连续传代后变异情况,根据已发表的CSFV E0基因序列(AF092448.2),设计合成引物,以在ST细胞中连续传代培养的CSFV石门株细胞毒总RNA为模板,每5代通过RT-PCR方法扩增病毒的E0基因,测定其核苷酸序列和氨基酸序列,用DNA Star软件分析比较原代、5代、10代、15代、20代、25代接毒细胞中CSFV E0基因的变异性。结果显示,接种病毒第10代的病毒E0基因在630核苷酸位点出现变异,第15代病毒在632核苷酸位点出现变异。本研究通过分子生物学方法发现CSFV Shimen株在ST细胞连续传代过程中E0基因能保持遗传的稳定性。

Variation Analysis of E0 Gene of Classical Swine Fever Virus Shimen Strain in Continuously Passed ST Cells

In order to study genetic variation of classical swine fever virus Shimen strains,the primers of CS-FV E0 gene of Shimen strain were designed and synthesized according to the published sequence (AF092448.2).The total RNA of classical swine fever virus Shimen strain continuouly passed in ST cells used as a template,and E0 gene was amplified by RT-PCR every five passages of cultured swine fever vi-rus.E0 nucleotide and amino acid sequences were determined.Variabilities of primary,5,10,15,20 and 25 passage viruses were analyzed by using DNA Star software.The results showed that variation appeared in 630 nucleotide point of 10 passage virus,and 632 nucleotide point of 15 passage virus.By molecular biologi-cal methods,this research revealed that the E0 genes of CSFV Shimen strain were genetically stable after passages in the ST cells.