重组胸腺素α_1的克隆、表达与纯化  

Cloning expression and purification of recombinant thymosin alpha-1

作　　者：宫照龙[1] 蒋作君[1] 徐义辉[1] 沈继龙[1] 董炜[1] 段招军[1] 

机构地区：[1]安徽医科大学病原生物学教研室,合肥230032

出　　处：《安徽医科大学学报》2004年第2期110-113,共4页Acta Universitatis Medicinalis Anhui

摘　　要：目的　利用基因工程方法表达重组胸腺素α1(Tα1) ,并进行纯化、鉴定。方法　使用大肠杆菌偏爱密码子 ,人工合成Ref蛋白 (T7噬菌体蛋白 )和Tα1的融合基因 ,并克隆在表达载体 pBV2 2 0中 ,转化E .coliDH5α ,温度诱导表达 ,表达的重组蛋白Ref Tα1经CM SepharoseFF阳离子柱进行纯化 ,Western blot进行鉴定。结果　获得了纯化的重组蛋白Ref Tα1,相对分子质量约为 15 0 0 0。结论　用融合基因的方法表达小分子多肽是一种可行的方法 ,Ref Tα1的成功表达为进一步研究其生物学活性奠定了基础。Objective To express recombinant thymosin alpha-1 (Tα_1) with gene engineering method and purify and identify the fusion protein.Methods The fusion gene composed of Ref(a sort of T7 phage protein)and Tα_1 gene was synthesized,and the gene was cloned into expression vector pBV220, E.coliDH5α was transformed with recombinant plasmid and induced by temperature.The expressed fusion protein could be purified by CM-Sepharose Fast Flow cation exchange chromatography and identified by Western-blot.Results The purified fusion protein,Ref-Tα_1 was obtained. The molecular weight was about 15 000 .Conclusion To express short peptide,the method of expressing fusion gene is practical.The successful expression of Ref-Tα_1 has established a foundation for biological activity research.

关 键 词：重组胸腺素α1 克隆 表达 纯化 遗传学 基因融合 基因表达 

分 类 号：R394[医药卫生—医学遗传学]