HIV-1 <em>Tat</em>基因重组原核表达载体的构建及融合蛋白的表达纯化与功能鉴定

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[Abstract] Objective: To construct the prokaryotic expression vector carrying HIV-1 Tat gene and to purify the fusion protein in E.coli. Methods: The DNA fragment of the Tat gene from pcDNA3-1（+）／Tat101 was cloned into the prokaryotic expression vector pET-28a(+),named pET-28a(+)Tat. Then the recombinant vector was transformed into E.coli BL21 (DE3). The expression of the histidinetagged (His-Tag) fusion protein was induced with isopropyl-β-D-thiogalactopyranoside (IPTG). The fusion protein was detected and purified by Western blot and immobilized Ni2+absorption chromatographic column, respectively. Results: The prokaryotic expression vector carrying Tat gene was successfully constructed. The expression of the His-Tag fusion protein in E.coli BL21 (DE3) was detectable. Finally, the fusion protein with the relative molecular weight of 18×103 was gained after purified using the affinity chromatographic column. Conclusion: Recombinant Tat gene can be expressed in E. coli BL21 (DE3) and the fusion protein obtained can be functionally active.

Received: 16 May 2013

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http://zzs.ujs.edu.cn/xbyxb/EN/ OR http://zzs.ujs.edu.cn/xbyxb/EN/Y2013/V23/I4/308

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