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《Journal of Hubei University(Natural Science)》 2018-01
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Expression,purification and activity analysis of human rhinovirus 3C protease in Escherichia coli

ZHOU Yu;XU Hu;YU Chan;ZHAI Chao;Bio-Resource Green Transformation Collaborative Innovation Center,College of Life Sciences,Hubei University;  
Human rhinovirus 3 C protease has high specificity to substrate,and it remains high activity under low temperature. This report was aim to heterologous expression of human rhinovirus 3 C protease in Escherichia coli expression system with MBP tag to improve its solubility. The coding sequence of human rhinovirus 3 C protease was cloned into vector pRK792 and the recombinant plasmid was transformed into Escherichia coli BL21( DE3), followed by inducing with IPTG to gain MBP-LEVLFQGP-6 x His-human rhinovirus 3 C protease fusion protein. The target protein was purified with Ni-column. The purified human rhinovirus 3 C protease was used to digest MBP-LEVLFQGP-6 x His-xylanase. Xylanase with N-terminal 6 x His tag was detected its activity. The results show that human rhinovirus 3 C protease was expressed in Escherichia coli successfully. It recognized and cleaved between MBP tag and 6 x His tag,leading to human rhinovirus 3 C protease with only 6 x His tag. Then human rhinovirus 3 C protease was used to remove MBP tag,obtained active xylanase. In conclusion,the MBP tag increased the soluble expression of human rhinovirus 3 C protease,and the tag didn 't affect the activity of human rhinovirus 3 C protease,which was an advantage during purification and removed tag in the low temperature of unstable proteins.
【Fund】: 中国农业科学院油料作物研究所油料脂质化学与营养湖北省重点实验室开放课题
【CateGory Index】: Q78;R373
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