A study of recombinant protective H.pylori antigens

World Journal of Gastroenterology : WJG
Zheng JiangPi-Long Wang

Abstract

To construct a recombinant vector which can express M (r)26000 outer membrane protein (OMP) from Helicobacter pylori (Hp), and to obtain the vaccine protecting against Hp infection and a diagnostic reagent kit quickly detecting Hp infection. The gene encoding the structural M(r)26000 outer membrane protein of Hp was amplified from Hp chromosomal DNA by PCR, and inserted in the prokaryotic expression vector pET32a (+), which was transformed into the Top10 E. coli strain. Recombinant vector was selected, identified and transformed into BL-21(DE3) E. coli strain. The recombinant fusion proteins were expressed. The antigenicity of recombinant protein was studied by ELISA or immunoblotting and immunized Balb/c mice. The gene of M(r)26000 OMP was amplified to be 594 base pairs, 1.1% of the cloned genes was mutated and 1.51% of amino acid residues was changed, but there was homogeneity between them. The recombinant fusion protein encoded objective polypeptides of 198 amino acid residues, corresponding to calculated molecular masses of M (r)26000. The level of soluble expression products was about 38.96% of the total cell protein. After purification by Ni-NTA agarose resin columniation, the purity of objective protein became about 90%....Continue Reading

References

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Citations

Apr 30, 2003·World Journal of Gastroenterology : WJG·Yang BaiDian-Yuan Zhou
May 18, 2013·Asian Pacific Journal of Cancer Prevention : APJCP·Akbar KhalilpourRahmah Noordin
Jul 11, 2003·World Journal of Gastroenterology : WJG·Ya-Fei MaoShu-Ping Li

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