苏云金芽孢杆菌晶体蛋白的制备方法

Extraction of Crystal Protein from Bacillus thuringiensis

采用了2种液体培养基、2种培养条件对3种苏云金芽孢杆菌进行培养。镜检观察当90%以上的芽孢脱落时处理菌体,经碱液裂解后比较等电点沉淀和高速离心方法提取Bt蛋白的收率和纯度。实验得出,3种菌体在1/2NB培养基,28℃,200r/min培养3d,蛋白产量均较高,其中HD-1型菌株高达107.48mg/L。高速离心得到的蛋白收率低于等电点沉淀法,但其纯度较高,满足作为抗原的条件,为ELISA检测转基因食品中Bt蛋白奠定了基础。

Three species of Bacillus thuringiensis were cultured at two conditions by two kinds of medium. The thallus was initially treated with lysis when 90% of spores left cells, and then was treated by isoelectric point deposition and high-speed centrifugation respectively. Result indicated that the yield of Bt protein was very high when the three species were fermented in 1/2NB medium,28℃ ,200 r/min for 3 days, especially for HD-1 ,amount to 107.48mg/L. Although the quantity of Bt protein from high-speed centrifugation was lower than that from isoelectric point deposition, its purity was higher, which made it as antigen for the development of polyclonal antibodies and for the application of detecting transgenic modified Bt protein.