柯萨奇病毒B组3型中国分离株VP1基因真核表达系统的克隆  

Clone of eukaryotic expression system for VP1 gene of Coxsackievirus group B type 3 Chinese strain

作　　者：田野[1,2,3,4] 钟照华[1,2,3,4] 赵铁强[1,2,3,4] 黄建林 安毅 唐伟[1,2,3,4] 沈景霞[1,2,3,4] 钟学宽 孟繁超[1,2,3,4] 黄永麟 鲁凤民[1,2,3,4] 叶鸿瑁[1,2,3,4] 

机构地区：[1]哈尔滨医科大学第一临床医学院心内科 [2]哈尔滨医科大学基础医学院 [3]哈尔滨工业大学 [4]北京医科大学第三临床医学院

出　　处：《中国地方病学杂志》1999年第3期169-172,共4页Chinese Jouranl of Endemiology

基　　金：国家自然科学基金;黑龙江省自然科学基金

摘　　要：目的探讨构建柯萨奇病毒Ｂ组３型（ＣＶＢ３）基因疫苗的可行性。方法应用逆转录ＰＣＲ技术扩增ＣＶＢ３中国分离株ＶＰ１基因，通过ＴＡ克隆法构建ｐＣＲ２．１－ＣＶＢ３ＶＰ１，将ＣＶＢ３ＶＰ１亚克隆至真核表达载体ｐＣＥＰ４，构建真核表达系统ｐＣＥＰ４－ＣＶＢ３ＶＰ１，并进行酶切和测序鉴定。结果发现ＣＶＢ３的中国分离株与Ｎａｎｃｙ株的ＶＰ１基因基本一致，均编码２９３个氨基酸，其中有５９处核苷酸存在变异，但仅改变了１０处氨基酸编码，证实克隆的片段为ＣＶＢ３ＶＰ１基因，表达载体为ｐＣＥＰ４。结论实验成功地构建了ＣＶＢ３中国分离株的真核表达系统ｐＣＥＰ４－ＣＶＢ３ＶＰ１。Objective To construct the gene vaccine of Coxsackievirus group B type 3(CVB3).Methods The VP1 gene of CVB3 Chinese strain was amplified by means of reverse transcript polymerase chain reaction (RT-PCR) and pCR2.1-CVB3VP1 was constructed by TA cloning strategy.The newly eukaryotic expression system pCEP4-CVB3VP1 was constructed by subcloning,and confirmed by restrict analysis and sequencing.Results The VP1 gene of CVB3 Chinese strain has 59 bases differences from that of CVB 3 Nancy strain,some of them cause only 10 amino acid changes and others are nonsense mutations.Conclusions Our results may be useful for research of CVB3 immunological characteristics.

关 键 词：VP1基因 真核表达系统 柯萨奇B病毒 克隆 

分 类 号：R373.23[医药卫生—病原生物学]