Human Helicobacter pylori HspA subunit gene cloning and sequence analysis of.docVIP

 PAGE \* MERGEFORMAT 12 Human Helicobacter pylori HspA subunit gene cloning and sequence analysis of Author: Wu Sheng-Wei, Wu Sheng-yu, Yuan Tian-hong, Wang Mingyong, Wu Chenglong [Abstract] Objective: To obtain the Helicobacter pylori (Hp) heat-shock protein A (HspA) subunit genes and nucleotide sequence analysis and homology comparison. Methods: H. pylori chromosome to obtain DNA, PCR amplification to obtain HspA gene be directed into pGEX-4T-1 prokaryotic expression vector for nucleotide sequence analysis. Results: After enzyme digestion and sequencing confirmed that Hp HspA the success of gene cloning, DNA sequence published in Genbank sequence comparison of 13 bases are different, the homology of 96%; by the base sequence encoding amino acid residues presumed sequence of no variation, homology of 100%. Conclusion: Hp HspA successfully cloned into the pGEX 4T-1 prokaryotic expression vector for the expression of HspA and research laid the experimental basis. [Keywords:] Helicobacter pylori; HspA subunit; cloning, biotechnology; gene sequence [Abstract] Objective: To get heat shock protein A (HspA) subunit gene of Helicobacter pylori, analyze its DNA sequence, and compare its homology. Methods: Chromosome DNA of H.pylori was extracted and the HspA gene was amplified with Polymerase Chain Reaction (PCR). The obtained DNA fragment was cloned into prokaryotic expression vector pGEX-4T-1 and sequenced. Results: HspA gene was cloned successfully which was demonstrated by enzyme digesting and gene sequencing methods. Comparing with DNA sequences reported in Genbank, nucleotide sequence of the obtained DNA fragment was 96% identical with that of HspA gene with a variation of 13 base pairs. The homology in putative amino acids was 100%. Conclusions: HspA gene has been Cloned into prokaryotic expression vector pGEX 4T-1 successfully, which makes an experimental foundation for its expression and associated research. [Keywords:] Helicobacter pylori; HspA subunit;