Abstract:

Objective:To clone MOMP gene from genomic DNA of Chlamydia trachomatis serotype E and to construct prokaryotic expression plasmid of pGEX/MOMP,and achieve the fussion expression in the Bacterium coli(BL-21).This has lay the foundation for future Ct vaccine research.Methods:MOMP gene of Ct serotype E was amplified by polymerase chain reaction(PCR) and the fragment was cloned into the vector pGEX.The positive recombinant was transformed into Bacterium coli(BL-21),and it was identified by enzyme digestion,PCR amplification and sequencing.Then it was induced to expression and identified by SDS-PAGE and Western-blotting,and it was purified. Result:About 1.2kb MOMP gene was successfully isolated.The DNA sequence of MOMP was found to be the same as the nucleotide sequence published by GenBank; The molecular weight of expression product was 66kd ,which conformity to expectancy molecular weight.And I got a quantity of purified protein.Conclusion:The prokaryotic expression vector pGEX/MOMP was constructed successfully,and it was expressed in Bacterium coli(BL-21),and the protein was purified successfully.