婴幼儿轮状病毒VP7基因表达产物生物学活性研究  

Study on the bioactivities of Group A rotavirus VP7 gene expression product

作　　者：吴景华[1] 王海欣[1] 赵杰[1] 董爱英[1] 

机构地区：[1]华北煤炭医学院附属医院,河北唐山063000

出　　处：《山东医药》2009年第31期28-30,共3页Shandong Medical Journal

基　　金：唐山市科技局课题项目(08130227C)

摘　　要：目的诱导并分析克隆婴幼儿轮状病毒VP7基因表达及其生物学活性,为进一步构建轮状病毒基因工程疫苗奠定基础。方法扩增与纯化VP7基因,并与pEDM27/5载体进行连接,转化乳酸杆菌,乳糖进行诱导表达,用鼠抗Vp7蛋白单克隆抗体特异性结合表达的Vp7蛋白,并测定其生物学活性。结果成功扩增婴幼儿轮状病毒VP7基因,并转化pEDM27/5载体,通过蓝白斑筛选DNA阳性重组子,转化感受态乳酸杆菌,SDS-PAGE电泳可见乳糖诱导后一条分子量约为28kD的蛋白带高效表达,并能与抗婴幼儿轮状病毒VP7蛋白特异结合。结论重组乳酸杆菌表达的VP7蛋白具有良好的生物学活性;可用于研制开发轮状病毒基因工程疫苗。Objective To induce and analyze the cloned Group A rotavirus VP7 gene expression and its bioactivities, so as to lay a foundation for the construction of genetic engineering vaccine against rotavirus. Methods The VP7 gene fragment was amplified and purified, then combined with pEDM27/5 and transformed into lacotobacillus . After induced by lactose, the VP7 protein was combined specifically with mouse moneclonal antibody, and the bioactivities of which was determined. Results The VP7 gene was amplified and transformed into pEDM27/5 successfully. DNA（ ＋ ） recombinant was selected by blue and white plague, and the lacotobacillus were transformed , a protein chalaza of about 28 kD was found by SDS-PAGE after the induction of lactose, and which could be combined specifically with the VP7 protein against rotavirus. Conclusions VP7 protein of restructuring lacotobacillus has good bioactivities;this can lay a foundation for the further development of genetic engineering vaccine against rotavirus.

关 键 词：婴幼儿轮状病毒 pEDM27/5载体 乳酸杆菌 基因转化 

分 类 号：R373.2[医药卫生—病原生物学]