改良型TAT—VP3融合基因原核表达载体的构建及表达  

Prokaryotic Expression Vector Construction and Expression of Improved Fusion Gene TAT- VP3

作　　者：张启波[1] 赵庆华[2] 詹辉[1] 颜汝平[1] 杨峻峰[1] 李俊[3] 王剑松[1] 

机构地区：[1]昆明医学院第二附属医院泌尿外科,云南昆明650101 [2]昆明医学院第二附属医院妇科,云南昆明650101 [3]昆明医学院第一附属医院肾内科,云南昆明650032

出　　处：《医学新知》2010年第6期535-537,共3页New Medicine

基　　金：国家自然科学基金（30760251）

摘　　要：目的 构建改良型TAT-VP3融合基因原核表达载体,并表达目的 蛋白.方法 采用PCR法设计和扩增改良型TAT-VP3融合基因,克隆至表达载体pThioHisA中,构建非融合表达载体,转化大肠杆菌BL21（DE3）plyS,经IPTG诱导表达,15%SDS-PAGE鉴定.结果 成功构建了改良型TAT-VP3融合基因的原核表达载体,经酶切、测序鉴定,证明构建正确.经SDS-PAGE鉴定可见相对分子质量16400的特异性目的 条带,37℃诱导8h的蛋白表达量最高,且多数以包涵体形式存在.结论 已成功构建改良型TAT-VP3融合基因原核表达载体,并表达目的 蛋白,为进一步的蛋白制备和应用研究奠定了基础.Objective To construct prokaryotic expression vector for improved TAT - VP3 fusion gene and express the target proteins. Methods We designed and amplified improved TAT - VP3 fusion gene by PCR and cloned into vector pThioHisA and transformed the constructed recombinant plasmid to E. coli BI21 （DE3）plyS for expression under induction of IPTG and identified the expressed product by 15% SDS - PAGE. Results The prokaryotic expression vector for improved TAT - VP3 fusion gene was constructed successfully,which was corroborated by restriction analysis and sequencing, with relative molecular masses of 16 400 was expressed. The expressed product mainly existed in a form of inclusion body. The expression level reached a peak value 8h after induction. Conclusion The prokaryotic expression vector for improved TAT - VP3 fusion gene was constructed successfully, and the target protein was expressed,which laid a foundation of further study on protein manufacturing and application.

关 键 词：改良型TAT—VP3融合基因 载体构建 表达 

分 类 号：Q782[生物学—分子生物学]