Helicobacter pylori ahpC gene expression and the original.docVIP

 PAGE \* MERGEFORMAT 12 Helicobacter pylori ahpC gene expression and the original Of: Li Yanqing, Duan Guang-Cai, Song Chunhua, Cheung Kin-kwong, WANG Shu-ling, Zhang Weidong, Xi Garden [Abstract] Objective To construct the H. pylori (Helicobacter pylori, Hp) ahpC genes in prokaryotic expression system, expression and purification of Ahp protein. Methods PCR method from the Chinese isolates MEL-Hp27 chromosomal ahpC gene was amplified DNA fragment gene inserted into expression vector pET-30a construct the recombinant plasmid pET30a-ahpC. recombinant plasmid was transformed into E. coli after DNA sequencing E.coli BL21 (DE3), IPTG induction. with nickel ion affinity chromatography protein and used SDSidentification. Results PCR amplification of the ahpC gene length of 594bp, by enzyme digestion and sequencing analysis of the gene inserted into the vector with GenBank published sequence similarity of 99%, SDSrevealed that IPTG induced by the molecular mass The purpose for the 23ku protein, and the output is high. Conclusion The successful construction of the ahpC expression vector pET30a-ahpC, and expressed in E. coli. [Keywords:] Helicobacter pylori, ahpC gene, cloning, gene expression ChinaABSTRACT: Objective To construct a prokaryotic expression system of ahpC gene of Helicobacter pylori. Methods The ahpC gene was amplified from Hp chromosomal DNA by PCR technique and cloned into the expression vector pET-30a. The recombinant vector pET30a-ahpC was identified by DNA sequencing and transformed to E.coli BL21 (DE3) for expression under induction by IPTG. The expression product was analyzed by SDS. Results PCR product showed that ahpC gene consisted of 594bp. The gene fragment that was inserted into the recombinant vector was identified to GenBank for 99%. SDSshowed that the induced protein was expressed highly in the host bacterium. Conclusion A prokaryotic high-expression system for ahpC gene has been successfully constructed. It can highly express