蓝藻抗病毒蛋白-N 衍生物的基因克隆、中试发酵与纯化

Objective：The cDNA of SUMO-L10-CVN gene was cloned and expressed in prokaryotic cells, and the pilot fermentation was completed. Methods:Through molecular design and optimization, a 10-amino acids flexible peptide (GGGGS)2 was linked to the N-terminus of CVN. NdeⅠand BamH I was chosen as the restriction enzyme site of sense primer and antisense primer ,thus the cDNA of SUMO-L10-CVN was cloned into plasmid pET-3c. SUMO-L10-CVN was expressed in a soluble form by SUMO fusion expression system in the cytoplasm of E. coli BL21. The recombinant L10-CVN was purified to homogeneity by Ni-NTA affinity chromatography after SUMO protease cleavage. Results:SUMO-L10-CVN was highly expressed in a soluble form in E coli BL21 under the induction of IPTG (20℃, 0.5 mmol/L).The fusion protein was expressed up to 25 % of the total protein. Conclusion: The fusion protein of SUMO-L10CVN was successfully expressed and purified in E. coli BL21(DE3).This culture method and its optimization of fermentation conditions laid a foundation of large-scale preparation of this recombinant protein.