Construction of hpaA gene from a clinical isolate of Helicobacter pylori and identification of fusion protein.

ABSTRACT:

Aim

To clone hpaA gene from a clinical strain of Helicobacter pylori and to construct the expression vector of the gene and to identify immunity of the fusion protein.

Methods

The hpaA gene from a clinical isolate Y06 of H.pylori was amplified by high fidelity PCR. The nucleotide sequence of the target DNA amplification fragment was sequenced after T-A cloning. The recombinant expression vector inserted with hpaA gene was constructed. The expression of HpaA fusion protein in E.coli BL21DE3 induced by IPTG at different dosages was examined by SDS-PAGE. Western blot with commercial antibody against whole cell of H.pylori as well as immunodiffusion assay with self-prepared rabbit antiserum against HpaA fusion protein were applied to determine immunity of the fusion protein. ELI

SUBMITTER: Mao YF 

PROVIDER: S-EPMC4615498 | biostudies-literature | 2003 Jul

REPOSITORIES: biostudies-literature