改良型TAT-VP3融合蛋白的基因合成、原核表达载体构建及表达的实验研究

Experimental Study on the Gene Synthesis,Prokaryotic Expression Vector Construction and Expression of Improved Fusion Protein TAT-VP3

目的研究改良型TAT-VP3融合蛋白的基因合成、原核表达载体的构建及其原核表达.方法首先通过基因拼接法合成VP3基因并委托生物技术公司合成改良型TAT基因.然后于原核表达载体pGEX-6p-1上构建改良型TAT-VP3融合蛋白的基因.最后将构建好的原核表达载体转导入基因工程菌DH-5α中并诱导其表达.结果成功合成了VP3基因,构建了改良型TAT-VP3融合蛋白的原核表达载体,并经基因测序证明构建无误;成功的在基因工程菌DH-5α中诱导了改良型TAT-VP3融合蛋白的表达.结论改良型TAT-VP3融合蛋白的基因合成、原核表达载体的构建及其原核表达是可行的,为进一步的蛋白制备及活性研究打下了基础.

Objective To explore the gene synthesis of fusion protein TAT -VP3, construction of the prokaryotic expression vector pGEX-6p-I-TAT/VP3 and the expression of this fusion protein in E.coli DH-5a. Methods First, the gene of VP3 was synthesized through gene splicing. At the same time, the sequence of improved TAT was designed and then a special biological engineer company was asked to compose it. The second step was to construct the prokaryotic expression vector pGEX-6p-I-TAT/VP3. At last, this prokaryotic expression vector was transplanted into E. coli DH-5a, then the expression of the fusion protein TAT /VP3 was induced. Results The gene of VP3 was synthesized and the prokaryotic expression vector pGEX-6p-I-TAT/VP3 was constructed successfully, Which was corroborated by the analysis of gene sequence. The fusion protein of improved TAT -VP3 in the E. coli DH-5a expressed successfully. Conclusions Gene synthesis, prokaryotic expression vector construction and expression of improved TAT-VP3 are feasible. The research may provide a foundation for the future work such as protein manufacturing and functional research of thisfusion protein.