摘要
本实验室使霍乱肠毒素(CT)B亚单位基因在大肠杆菌中获得表达,并能分泌到胞外。将该菌株培养物上清经过超滤浓缩后,用偶联上霍乱毒素IgG的CNBr-Sepharose 4B进行亲和层析,得到表达产物霍乱毒素B亚单位纯蛋白。经PA-GE、HPLC及琼脂免疫扩散等方法鉴定证明该蛋白与天然霍乱肠毒素B亚单位完全相同。
The recombinant plasmid pMM2 contained cholera enterotoxin B sub-unit gene CT-B were expressed in E.coli strain. Cholera toxin B subunit was purified by immunoaffinity chromatography from culture supernatant of the strain by using CNBr-Sepharose 4B coupled with CT-IgG as affinity medium. The purified product is homogeneous and identical with the B subunit from wild V.cholera strain as shown by SDS-PAGE, HPLC and immunodiffusion.
关键词
霍乱肠毒素
蛋白纯化
亲和层析
cholera toxin, purification of protein, affinity chromatography
