This article reviews several cell breakage methods, such as extraction buffer method, ultrasonic disruption, repetitive freezing and thawing, bugbuster and high-pressure cracker. Compare the periplasmic protein we get from each mehod, in aspects of weight and activity, by using SDS-PAGE electrophoresis, BCA Protein Assay Kit, and ELISA. The conclusions are as follows: in a small scale laboratory , ultrasonic disruption is the most efficacious method, and the optimal condition is putout power 250W, radiation/intermission time 4s/5s, total working times is 140, and the amount of cell buffer 5mL/g with ice bath. Besides, for the large-scale periplasmic protein preparation, the optimal method is high-pressure cracker in 850Pa, crushing twice.
陈璩1 罗勤2 夏亮2 王明蓉1. 大肠埃希菌周质抗体细胞释放工艺方法研究[J]. J4, 2010, 35(4): 270-.
Chen Qu, Luo Qin, Xia Liang and Wang Ming-rong. Research for extraction of periplasmic proteins in Escherichia coli. J4, 2010, 35(4): 270-.