Expression And Immunoreactivity Analysis Of Hspa Gene From Helicobacter Pylori In Lactococcus Lactis

Objective:To construct a recombinant plasmid containing gene encoding heat shock protein A (HspA) from Helicobacter pylori (Hp) and express it in Lactococcus lactis, and to explore the immunoreactivity.Methods:The hspA gene was amplified from genome DNA of Hp NCTC11637 strain by polymerase chain reaction (PCR), and then cloned in the E. coli-L.lactis shuttle vector pMG36e and transformed into E.coli DH5É‘. The positive recombinant plasmid was named pMG36e/hspA. and then electroporation was performed to transformed the recombinant plasmid into L.lactis MG1363. The expression of HspA protein was detected by Western blotting.Results :We have constructed the pMG36e/hspA by cloning and recombinant DNA technique. The Western blotting analysis showed that the relative molecular mass (Mr) of expression product was about 13kDa, and could be recognized by anti-HspA serum produced by oral administration of the L.lactis transformant into mice, suggesting that the recombinant HspA protein had good immunoreactivity.Conclusion:The recombinant L.lactis MG1363 expressing HspA protein of Hp was constructed successfully. The results lay the foundation for the further research of Hp vaccine.