改良型TAT-VP3融合蛋白的表达、纯化及鉴定  

Expression,Purification and Identification of Improved TAT-VP3 Fusion Protein

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

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]昆明医学院第一附属医院肾内科,云南昆明650032

出　　处：《昆明医学院学报》2010年第9期4-6,26,共4页Journal of Kunming Medical College

基　　金：国家自然科学基金资助项目(30760251)

摘　　要：目的表达改良型TAT-VP3融合蛋白,并对其进行纯化.方法利用IPTG温控诱导改良型TAT-VP3融合蛋白表达,并经GST亲和层析纯化,目的蛋白进行SDS-PAGE和Western bolt鉴定.结果改良型TAT-VP3融合蛋白经温控诱导后得到稳定表达,经SDS-PAGE鉴定可见相对分子质量42 kD的特异性目的条带,经GST亲和层析纯化后,得到了纯度较高的目的蛋白.Western bolt验证了表达蛋白的特异性.结论已成功表达及纯化出改良型TAT-VP3融合蛋白,为进一步研究其免疫原性和抗肿瘤活性奠定了基础.Objective To express and purify improved TAT-VP3 fusion protein.Methods The improved TAT-VP3 fusion protein was expressed under the induction of temperature by IPTG.The expressed product was purified by GST affinity chromatography and identified by SDS-PAGE and Western blot.Results The improved TAT-VP3 fusion protein was stably expressed under the induction of temperature.The specific band with relative molecular masses of 42000 was identified by SDS-PAGE.The expressed product was purified successfully by GST affinity chromatography and showed good specipicity as proved by Western blot.Conclusion The improved TAT-VP3 fusion protein has been successfully expressed and purified,which lay a foundation for further study on its immunogenicity and antitumor activity.

关 键 词：改良型TAT-VP3融合蛋白 表达 纯化 鉴定 

分 类 号：R996.3[医药卫生—毒理学]