Gene cloning,expression and biological function of hLN?4LG1 / 第三军医大学学报

ABSTRACT

Objective To clone and express human laminin alpha4 chain LG1 module(hLN?4LG1)protein holding biological activity.Methods The cDNA encoding hLN?4LG1 was amplified by RT-PCR,then inserted into pMD-18T vector and sequenced.The hLN?4LG1 cDNA fragment was subcloned into pET-28a vector and expressed in BL21(DE3) strain.The hLN?4LG1 protein was detected by Western blotting and purified by Ni-NTA column.The biological activity of target protein was detected through cell adhesion and expansion test.Results The cDNA fragment of hLN?4LG1 was cloned successfully.While BL21(DE3)/pET-28a-LG1 bacterium was induced with IPTG,a new protein band with a relative molecular weight of 26 000 was shown.Purified hLN?4LG1 protein could promote the expansion and adhesion of A549 cells obviously,as compared with the control group.Conclusion The hLN?4LG1 was cloned and expressed successfully,on which the cells can adhere and expand.