大肠杆菌生产基因重组草鱼生长激素的分离纯化

Isolation and Purification of Recombinant Grass Carp Growth Hormone Produced in Escherichia coli

草鱼生长激素cDNA在大肠杆菌细胞中表达后，重组草鱼生长激素在胞内形成不溶性的包含体．这种包含体在洗涤、变性、复性和C12反相层析等步骤后，产物再经快速蛋白液相色谱，重组草鱼生长激素在SDS-聚丙烯酰凝胶电泳中显示均一性，纯度可达90％放射免疫分析表明：重组草鱼生长激素具有草鱼生长激素的免疫活住．分离纯化产率约为1L细菌培养液1～2m重组草鱼生长激素．

Cloned cDNA of grass carn growth hormone (rgGH) was expressed in Escherichia coli cells and the rgGH was accumulated as insoluble inclusion theies. The inclusion bodies were washed and denatured. After renaturation the rgGH was purified by C15 revcrse phase chromatography and fast protein liquid chromatography (FPLC) to 90% purity. The results of radioimmunoassay show that the rgGH possesses immunoreatctivity of growth hormone. The final yield of the rgGH was 1 ~ 2 mg per liter of bacteria culture.