Cloning and expressing of Decorin gene / 西安交通大学学报(医学版)

ABSTRACT

ObjectiveTo study Decorin biological effects,Decorin gene cDNA was cloned and expressed in E. coli. MethodsThe Decorin gene obtained by PCR from T cell cDNA library and cloned into plasmid vector pUC19. The encoding sequence of Decorin in the pUC19 was confirmed by DNA sequencing using Sanger Dideoxy method, and then it was subcloned into expression plasmid vector pGEX-4T-1, the recombinant vector was indentified with BamHI and EcoRI in 1.2 ％ agarose gel electrophoresis. The expression of Decorin fusion protein with GST in E. coli JM109 was induced with IPTG and identified by SDS-PAGE. ResultsThe PCR amplified DNA fragment shared identical sequence with known Decorin gene reported in GenBank (Accession number AF138300). The SDS-PAGE analysis revealed that the expressed fusion protein MW was approximatelly 65.6KD and in soluble format. ConclusionThe cloned Decorin gene is correct and it was expressed in fusion protein with GST.