Establishment Of Prokaryotic Fusion Expression System With Human SUMO1/2/3/4-tag

Today the demands of scientific research and therapeutic drug production for large quantities of native protein has increasd exponentially.The gene expression technology is very good solved the problem.The technology including eukaryotic exprssion system and prokaryotic expression system.The prokaryotic expression system is most commonly used for easy operation and low cost. In the prokaryotic expression system inclusion body easy generation to purification of faults, scientists also designed fusion expression systems. In these fusion expression system,SUMO fusion system depending on its function fusion expression system to promote dissolve expression and high precision cutting and become a star of the limelight. We tried to get away from the four human proteins, looking for has the highest expression and promote soluble promoting effect of members as fusion tags to establish the prokaryotic fusion expression system.This study in 4 human function proteins as fusion tags to PET28a as a carrier skeleton, build four fusion expression vector pS1, pS2, pS3, and pS4. Then with P53 protein and GFP protein as fusion respectively target protein is inserted into the the 4 SUMO-tag expression vector and PET28a , and get 10 kinds of fusion expression vector(pS1-P53, pS2-P53, pS3-P53, pS4P53, pS1-GFP, pS2-GFP, pS3-GFP, pS4-GFP,p28-P53,p28-GFP). Put these fusion vector transformation expression strain——BL21-CodonPlus(DE3)-RI. For induction expression, and purify product. Meanwhile induction GST-SUMO1-SUMO1 protein. Its purification products as substrate and SENP1C for enzymatic reaction. Detection of active function of SUMO protease.Research found that SUMO1 as fusion to tag, P53 and GFP two for recombinant protein and expression and promote dissolve effect is the best. We induce expression SENP1C has good soluble. The product of purified with substrate GST-SUMO1-SUMO1 protein reaction. After cutting strip appears GST-SUMO1. This shows SENP1C with activity.