Construction and expression on prokaryotic expression vector of pfs gene together with recombinant protein purification

The pfs gene was amplified by polymerase chain reaction( PCR) using the DNA of Lactobacillus plantarum KLDS1.0391 as template in this study. The nucleotide sequence of the cloned DNA shared 99%homology with pfs gene from Lactobacillus plantarum JDM1 and WCFS1. The target gene was connected with expression vector p QE-30 and the recombinant plasmid p QE-30-pfs was transformed into E.coli M15. The recombinant expressed proteins induced by IPTG were analyzed and identified with SDS-PAGE. SDS-PAGE results showed that there was one main protein band with molecular weight 27.4ku in the total protein. The recombinant protein was purified using Ni-NTA Purification System and identified with SDS-PAGE. The results showed that the prokaryotic expression vector p QE-30-pfs was constructed successfully. Pfs protein was expressed successfully in E.coli M15. Recombinant protein Pfs was purified successfully using Ni-NTA Purification System. This results will be helpful for the synthesis of AI-2 in vitro.