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. 2004 Sep 1;10(17):2560-2.
doi: 10.3748/wjg.v10.i17.2560.

Cloning and expression and immunogenicity of Helicobacter pylori BabA2 gene

Yang Bai  1 , Ya-Li Zhang, Ye Chen, Jian-Feng Jin, Zhao-Shan Zhang, Dian-Yuan Zhou
Affiliations

Cloning and expression and immunogenicity of Helicobacter pylori BabA2 gene

Yang Bai et al. World J Gastroenterol. .

Abstract

Aim: To construct a recombinant strain which expresses BabA of Helicobacter pylori (H pylori) and to study the immunogenicity of BabA.

Methods: BabA2 DNA was amplified by PCR and inserted into the prokaryotie expression vector pET-22b (+) and expressed in the BL21 (DE3) E.coli strain. Furthermore, BabA immunogenicity was studied by animal test.

Results: DNA sequence analysis showed the sequence of BabA2 DNA was the same as the one published by GenBank. The BabA recombinant protein accounted for 34.8% of the total bacterial protein. The serum from H pylori infected patients and Balb/c miced immunized with BabA itself could recognize rBabA.

Conclusion: BabA recombinant protein may be an potential vaccine for control and treatment of H pylori infection.

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Figures

Figure 1
Figure 1
BabA2 DNA fragment amplified by PCR from Helicobacter pylori electrophoresed on 8 g/L agarose gel. Lane 1: Nucleotide marker; Lane 2: PCR products.
Figure 2
Figure 2
Identification of recombinant plasmid by restriction enzyme digestion. Lane 1: Nucleotide marker; Lane 2: PCR products; Lane 3: Recombinant plasmid/Not I; Lane 4: Recombinant plasmid/Not I and Nco I; Lane5: pET22b (+)/Not I.
Figure 3
Figure 3
The 100 g/L SDS-PAGE analysis of fusion protein expressed in BL21 (DE3). Lane 1: Molecular mass marker (67, 94) × 103; Lane 2: control strain BL21 (pET) before induction; Lane 3: control strain BL21 (pET) after 5-h induction with IPTG; Lane 4: BL21 (pET-BabA) cells before induction; Lane 5: BL21 (pET- BabA) cells after 3-h induction with IPTG; Lane 6: BL21 (pET- BabA) cells after 5-h induction with IPTG; Lane 7: BL21 (pET- BabA) cells periplasm protein after 3-h induction with IPTG; Lane 8: sonicated supernatant of BL21 (pET-BabA) cells after 3-h induction with IPTG; Lane 9: recombinant protein BabA purified.
See this image and copyright information in PMC

References

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