荷兰弱毒株凋亡素基因的克隆及真核表达载体的构建  

Clone of apoptin gene of Netherlands attenuated strain and construction of its eukaryotic expression vector

作　　者：于健[1] 韩国均[1] 王立冬 李梅[3] 赵岩[1] 

机构地区：[1]哈尔滨医科大学附属第一医院耳鼻喉科,黑龙江哈尔滨150001 [2]哈尔滨市第四医院,黑龙江哈尔滨150026 [3]哈尔滨市第一医院耳鼻喉科,黑龙江哈尔滨150010

出　　处：《哈尔滨医科大学学报》2011年第1期5-8,共4页Journal of Harbin Medical University

基　　金：黑龙江省教育厅科学技术研究项目(11531200)

摘　　要：目的克隆鸡贫血病毒荷兰弱毒株的凋亡素基因,比较来自不同毒株的凋亡素的分子结构差异。方法经过两次PCR,将含VP2、VP3 DNA片段粘端连接克隆到pUC119质粒中,再将VP3 DNA片段平端连接克隆到pUC119质粒中。结果成功克隆到荷兰弱毒株凋亡素基因,并构建了其真核表达载体。结论荷兰弱毒株的凋亡素与26P4等5个毒株有不同的蛋白质二级结构。Objective To clone the Netherlands attenuated strain of chicken anemia virus apoptin gene,a comparison of different strains of different molecular structure of Apoptin.Methods After two PCR,VP2,VP3 DNA fragment was cloned into pUC119 sticky end to connect plasmid,then VP3 DNA fragment was cloned into pUC119 flush end connections plasmid.Results The Netherlands avirulent apoptin gene was successfully cloned and its eukaryotic expression vector was constructed.Conclusion Netherlands attenuated strain of apoptin and 26P4 of five strains have different protein secondary structure.

关 键 词：鸡贫血病毒 凋亡素 聚合酶链反应 分子克隆 序列测定 

分 类 号：Q78[生物学—分子生物学] R373[医药卫生—病原生物学]