鼠源轮状病毒抗原基因的表达载体构建及农杆菌转化研究  

Construction of murine rotavirus antigen gene expression vector and transformation the recombinant plasmids into Agrobacterium tumefaciens

作　　者：李晋涛[1] 吴玉章[1] 费蕾[1] 唐艳[1] 邹丽云[1] 

机构地区：[1]第三军医大学基础医学部全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2002年第8期892-894,共3页Journal of Third Military Medical University

基　　金：国家自然科学基金资助项目 ( 397890 10 )

摘　　要：目的　构建表达鼠源轮状病毒抗原基因vp4的植物载体及农杆菌转化。方法　抗原基因vp4经PCR扩增后直接克隆到pUC T载体 ,双酶切目的片断和植物表达载体 ,以T4连接酶将目的片断与载体相连接 ,电转化方式将重组质粒转入农杆菌。结果　成功地将目的基因定向克隆到表达载体 ,并转入到农杆菌中。结论　pUC T载体可直接进行PCR扩增产物克隆 ,便于测序 ;电转化方式可以有效、快速地将大分子量质粒 (>10kb)Objective To construct plant expressing murine rotavirus antigen gene and transform it into Agrobacterium tumefaciens. Methods PCR product of vp4 gene was directly cloned into pUC T vector, and the vp4 DNA fragments were cut down by restriction enzymes and inserted into the plant expression vector. The recombinant plasmids were transformed into Agrobacterium tumefaciens by electroporation. Results Murine rotavirus antigen gene vp4 was successfully cloned into the plant expression vector PBI121 and the recombinant plasmids were efficiently transformed into the target bacterium. Conclusion pUC T vector is a functional vector. Electroporation is a quicky and efficient way to transform large weight molecular plasmids (>10 kb) into Agrobacterium tumefaciens .

关 键 词：抗原基因 农杆菌 轮状病毒 载体构建 电击转化 VP4基因 植物载体 小儿腹泻 轮状病毒疫苗 

分 类 号：R722.132[医药卫生—儿科]