维氏气单胞菌Cbl蛋白的原核表达及纯化

维氏气单胞菌(Aeromonas veronii)引起的细菌性疾病已威胁到鱼类养殖业的发展.Cbl(类CysB蛋白)是细菌硫代谢的重要调控因子之一.本研究选取pET-28a为Cbl的原核表达载体,以大肠杆菌BL21(DE3)为宿主菌,使用IPTG(Isopropyl-beta-D-thiogalactoside,异丙基...

Prokaryotic Expression and Purification of Cbl Protein from Aeromonas veronii

Bacterial diseases caused by Aeromonas veronii have threatened the development of fish production. And Cbl (CysB like protein) is one of the important regulatory factors of bacterial sulfur metabolism. Escherichia coli BL21 (DE3) was selected as the prokaryotic expression vector for Cbl, and culturedadded with IPTG (Isopropyl-beta-D-thiogalactoside) for culture to induce the expression of Cb1. The protein samples were run on SDS-PAGE gel electrophoresis to verify the protein bands of Cb1, and then the concentration of Cbl was determined with Boster’s BCA protein assay kit. The results showed that Cbl was successfully purified, and that the concentration of Cbl was up to 601.405 mg·L?1, which meets the requirements for subsequent experiments, when the induction time was 8h, the final IPTG concentration was 0.1 mmol·L?1, the induction temperature was 15 ℃ and the Imidazole concentration in the elution buffer was 100 mmol·L?1.