摘要
目的 :建立稳定表达丙型肝炎病毒 (HCV)核心蛋白的原核表达系统 ,获得高产量的纯化核心蛋白。方法 :应用多聚酶链反应 (PCR) ,以HCV -H株全长cDNA序列为模板 ,扩增获得核心区基因片段 ,克隆入原核表达载体pBVIL1,构建原核表达载体pBVIL1-C ,转化HB10 1宿主菌 ,通过温度诱导表达核心蛋白。结果 :扩增得到目的基因长度为 5 73bp ,构建pBVIL1-C表达载体 ,在HB10 1宿主菌中通过温度诱导获得稳定表达 ,表达蛋白占菌体总蛋白含量的 2 1% ,Western -Blot检测证实表达产物可与HCV患者阳性血清发生特异性结合反应。结论
Objective:To construct a steady expression system of HCV Core protein in E.coli.Methods:The cDNA sequence encoding HCV Core gene were amplified by PCR and inserted into plasmid pBVIL1,the resulting expression vector was transformed into HB101,a strain of E.coli and expressed the protein by temperature induction.Results:The length of amplified Core gene is 573bp,pBVIL1 produced Core protein with yields of 21% of total proteins.The protein produced was detected by SDS-PAGE,western blot.And the protein showed specific immunoreactivity with serum from hepatitis C patient in western blot assay.Conclusion:HCV Core gene can be expressed in E.coli and exhibited immunogenicity.
出处
《生物技术》
CAS
CSCD
2004年第4期4-6,共3页
Biotechnology
基金
陕西省自然科学基金 (2 0 0 2C2 - 0 1 )
国家农业科技成果转化资金重点项目 (0 2EFN2 1 60 0 573)
