Objective  To express

purify and identify human P311 protein

a candidate protein in hypertrophic scar formation.   Methods  The cDNA sequence of human P311 obtained by PCR was cloned into the prokaryotic expression vector pGEX-4T-1 containing glutathione-s-transferase (GST). pGEX-4T-P311 was transformed into E. coli BL21(DE3). The expression of GST-P311 fusion protein was induced by IPTG

identified by both SDSPAGE and Western blotting

and then purified with glutathionesepharose beads.   Results  The restriction endonuclease digestion and the sequencing of recombinant plasmid demonstrated that pGEX-4T-

P311 vector was successfully constructed. The relative molecular weight of expressed protein was about 34×10

3

which was as the same as the objective protein

and the corresponding protein was identified by the immunoblotting with antibody against GST.   Conclusion  pGEX-4T-P311 vector is correctly constructed

and the GST-P311 fusion protein is successfully expressed in E. coli BL21(DE3)and purified.