Molecular Cloning And Expression Of RgpA And RgpB Catalytic Domain Genes Of Porphyromonas Gingivalis

Objective:To clone the rgpA and rgpB catalytic domain genes of Porphyromonas gingivalis(Pg) and to construct their expression system via pET42a vector.Methods:The rgpA and rgpB catalytic domain genes were amplified by polymerase chain reaction(PCR) and T-A cloned from Pg ATCC 33277 strain.The nucleotides of the target DNA fragments were firstly sequenced and then inserted into the pET42a to construct expression vectors.The expressions of rgpA and rgpB catalytic domain proteins were induced by IPTG with different dosages in BL21DE3 E.coll.Results:The homology of nucleotides sequence of the cloned rgpA and rgpB catalytic domain gene fragments from ATCC 33277 strains were 98.9%and 99.0% compared with the reported sequences.The homology of the amino acid sequences of the cloned rgpA and rgpB catalytic domain gene fragments were 99.2%and 99.1% compared with the reported sequence.The expression output of RgpA and RgpB catalytic domain(rgpAcd,rgpBcd) genes in pET 42a-rgpAcd/rgpBcd-BL21DE3 system was approximately 60%of the total bacterial proteins.Conclusion:An expression system of Pg rgpA and rgpB catalytic domain genes with high efficiency was successfully established.This will be the premise for the study on their immunogenicity and biological functions.