致病性鸡大肠杆菌pilA和外膜蛋白C原核诱导表达条件的优化

Optimized Expression of pliA and ompC from Avian Pathogenic Escherichia coli

分别从温度、接菌量、诱导时机、IPTG浓度、诱导时间等条件对2种重组菌株BL21/pET-28a-pilA和BL21/pET-28a-ompC进行优化.得到最佳诱导条件:BL21/pET-28a-pilA菌株为40℃摇培,80μL的接菌量,160 r/min摇培2 h后加入终浓度为0.96 mmol/L的IPTG,再诱导表达4 h,得到最高特异蛋白表达量;BL21/pET-28a-ompC菌株为37℃培养,80μL的接菌量,160 r/min摇培2.5 h后加入终浓度为0.64 mmol/L的IPTG,再诱导表达6 h,得到最高特异蛋白表达量.

Type Ⅰ piliA of avian pathogenic E. coli was a very important pathogenic factor. E. coli outer membrane protein had nicer immunogenicity and it played an important role in maintaining the form of bacteria, metabolism and immunity. The optimum conditions of induction from the temperature, amount of bacteria, induction opportunity, IPTG concentration and induction time in BL21/pET-28a-pilA and BL21/pET-28a-ompC were explored. The best condition of BL21/pET-28a-pilA was 40 ℃,80 μL was effective amount,shaking 2 h and then pipette IPTG into cuvettetests to make a final concentration of 0.96 mmol/L, and another 4h induction. The best condition of BL21/pET-28a-ompC was 37 ℃ , 80 μL was effective amount, shaking 2.5 h and then pipette IPTG into cuvettetests to make a final concentration of 0.64 mmol/L, and another 6 h induction. Thus, the highest expression levels of specific proteins were obtained.