Li Yan, Ning Yun-shan, Long Min, Dong Wen-qi, Li Ming
Department of Tropical Diseases, School of Biotechnology, Southern Medical University, Guangzhou 510515, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2006 Jul;22(4):440-2, 446.
To construct the recombinant plasmid containing catalase (KatA) of Helicobacter pylori (Hp), analyze its nucleic acid sequence, express it in E. coli and study its antigenicity.
KatA fragments were amplified from Hp chromosomal DNA by PCR. Its T-A was cloned, sequenced and compared with other HP strains on the GenBank. Then the gene cloned into pGEX-4T-1 fusion expression vector was expressed in E. coli and purified by GST-affinity chromatography. The purified product was used to identify 29 stains of mouse anti Hp monoclonal antibodies and analyze antigenicity with serum of Hp-infected patients by Western blot.
KatA fragments were composed of 1,515 bp (GenBank No. DQ333889) and the nucleotide homology with other Hp strains on the GenBank was 96%-97%. 85 kDa of the recombinant KatA-pGEX-4T-1 was expressed in E. coli. 4 of 29 anti-Hp mouse monoclonal antibodies were against KatA. Western blot analysis proved that KatA was specifically recognized in the serum of Hp-infected patients.
The recombinant KatA has original antigenicity. It is of great value to clinical sero-diagnosis and vaccine study of Hp.
