Cloning, expression and purification of the human augmenter of liver regeneration (hALR) cDNA / 第二军医大学学报

ABSTRACT

Objective: To clone human augmenter of liver regeneration (hALR) cDNA, construct the recombinant expression vector, express and purify its product. Methods: The hALR cDNA was obtained by using RT PCR method with total RNA extracted from the fetal hepatic tissue. Then it was cloned into the pGEM T vector, and subcloned into expression vector pGEX 4T 3.After proved to be correct by sequencing, recombinant expression plasmid pGEX 4T 3(hALR) was transformed into E.coli BL21(DE3). The fusion protein GST hALR was produced by IPTG induction, isolated by affinity chromatography glutathione Sepharose 4B and cleaved by Thrombin. Results and Conclusion: Recombinant expression plasmid pGEX 4T 3(hALR) is constructed. The hALR is highly expressed in E.coli . The fusion protein in the plasma is resoluble. The procedure of purification and cleavage of fusion protein is easy and simple.