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• W77893844 abstract "AIM: To construct a recombinant vector containing gene encoding heat shock protein A with a Mr of 13 000 from human Helicobacter pylori (H pylori) and express it in E. coli BL21, and to explore the antigenicity. METHODS: The target gene was amplified from H pylori chromosome by PCR, and then inserted into the prokaryotic expression vector pET32a (+) digested by restrictive endonuclease enzymes of kpn , BamH I simultaneously. The recombinant vector was transformed and expressed in E.coli BL21.The antigenicity of recombinant fusion protein was analysed by Western blot. RESULTS: Enzyme digestion and sequencing analysis showed that the target gene has been inserted into the recombinant vector, but as compared with the gene reported by GenBank, 1.6 % of gene mutation and 1.6 % of amino acid residues change in H pylori occurred, respectively. SDS-PAGE analysis showed that the recombinant vector could be expressed in E.coli BL21, the relative molecular mass (Mr) of expressed product was 33×103, while Mr of protein expressed by pET32a (+) was about 20×103, and soluble fusion expression product accounted for 18.96 % of total bacterial protein. After purification with Ni+-NTA agarose resin, the purity of recombinant fusion protein was about 95 %. Western blot result showed that recombinant fusion protein could be recognized by anti-H pylori positive serum, suggesting that the protein had good antigenicity. CONCLUSION: The gene encoding H pylori heat shock protein A has been cloned and expressed successfully. The results lay the foundation for development of H pylori protein vaccine and a quick diagnostic kit for detection of H pylori infection. Jiang Z, Pu D, Huang AL, Tao XH, Wang PL. Cloning, expression and antigenic analysis of heat shock protein A gene of human Helicobacter pylori. Shijie Huaren Xiaohua Zazhi 2003;11(10):1480-1484" @default.
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• W77893844 date "2003-10-15" @default.
• W77893844 modified "2023-09-24" @default.
• W77893844 title "Cloning, expression and antigenic analysis of heat shock protein A gene of human Helicobacter pylori" @default.
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• W77893844 doi "https://doi.org/10.11569/wcjd.v11.i10.1480" @default.
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