Expression,purification and cleavage activity analysis of self-processed recombinant MBP-HRV 3C fusion protease in E.coli expression system / 国际检验医学杂志
International Journal of Laboratory Medicine
; (12): 2721-2722,2725, 2014.
Article
in Zh
| WPRIM
| ID: wpr-600277
Responsible library:
WPRO
ABSTRACT
Objective To obtain a novel tool-enzyme for genetic engineering with good solubility,strong specificity of enzyme digestion and maintaining the enzyme activity at low temperature by using E.coli expression system to express self-processed re-combinant MBP-HRV 3C fusion protease.Methods The cDNA encoding HRV 3C protease was cloned into pRSF-Duet vector.The recombinant plasmid was transferred into E.coli BL21 (DE3)for expression.HRV 3C protease was obtained through Nichol col-umn affinity purification.The cleavage activity of HRV 3C protease was determined by in vivo experiment.Results HRV 3C prote-ase was highly expressed in E.coli expression system,and the obtained HRV 3C protease could recognize and digest HRV 3C site. Conclusion A novel tool-enzyme for genetic engineering is obtained.
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WPRIM
Language:
Zh
Journal:
International Journal of Laboratory Medicine
Year:
2014
Type:
Article