猪乙脑病毒分离株CSF.XZ2D在BHK21细胞上的传代培养及E基因序列分析  

Sequence Analysis of the E Gene of JEV Isolate CSF.XZ-2D Passaged in BHK-21 Cell Line

作　　者：梁晓晓[1,2] 罗俊[2] 禹乐乐[2,3] 滕蔓[2] 胡博[2] 杨艳艳[2] 张改平[2] 钟秀会[1] 

机构地区：[1]河北农业大学动物科技学院,河北保定071000 [2]河南省农业科学院农业部动物免疫学重点实验室/河南省动物免疫学重点实验室,河南郑州450002 [3]郑州大学生物工程系,河南郑州450001

出　　处：《河南农业科学》2013年第5期153-157,共5页Journal of Henan Agricultural Sciences

基　　金：国家公益性行业(农业)科研专项(201203082-5)

摘　　要：为了解河南省规模化猪场日本乙型脑炎病毒(JEV)流行株的特征及其遗传基因的稳定性,将猪乙脑病毒分离株CSF.XZ-2D在BHK-21细胞上进行连续传代培养,并对其E基因的遗传稳定性进行了研究。结果表明,经连续传代25次后病毒E基因趋于稳定,氨基酸位点E271(E→V)和E278(V→L)传代后发生稳定点突变。与JEV毒力相关的部分位点如E107、E138、E176、E177和E315遗传稳定性较高,经连续传代后均未发生突变,与SA14-14-2相应位点完全一致。但是,另外一些位点的遗传稳定性较差,如E279和E312分别出现K→M→K和K→R→K→R的反复突变。这些突变是否与JEV的宿主细胞适应性及毒力变化相关有待进一步研究。To investigate the features of Japanese encephalitis virus（JEV） currently pravalent and the genetic stability in pig farm in Henan province,the genetic stability of E gene of JEV isolate CSF.XZ-2D passaged in BHK-21 cells（baby hamster kidney cell line） was studied.The results showed that the E gene became stable after 25 passages in BHK-21 cells.After serial subcultivation,two stable amino acid（aa） mutations happened at the sites E271（E→V） and E278（V→L）.No mutations were observed at part of the virulence correlated aa sites,such as E107,E138,E176,E177 and E315,same to those of vaccine strain SA14-14-2.However,mutations at the other two virulence correlated aa sites of E279（K→M→K） and E312（K→R→K→R） happened repeatedly.Whether these mutations are correlated with the virulence variation and adaptability of JEV to host cells needs to be further studied.

关 键 词：乙脑病毒 CSF XZ-2D BHK-21 传代培养 E基因 序列分析 

分 类 号：S852.651[农业科学—基础兽医学]