牛产肠毒素性大肠埃希菌菌蜕的制备及裂解条件的优化

Preparation of bacterial ghost of bovine enterotoxigenic E. coli and optimization of condition for pyrolysis

目的制备牛产肠毒素性大肠埃希菌菌蜕,并对裂解条件进行优化。方法将裂解质粒pHH43电转入牛肠产毒素性大肠埃希菌中,37℃培养至A_(600)值为0.6时,升温至42℃诱导裂解,制备菌蜕。同时对起始诱导A_(600)值(0.4、0.6、0.8、1.0、1.2)和培养基成分含量(2/3、3/4、4/5和正常含量)进行优化,并计算裂解效率。结果在起始诱导A_(600)值为0.6,培养基营养成分为正常含量的3/4时,裂解效率最高,达99.93%。结论成功制备了牛产肠毒素性大肠埃希菌菌蜕,并对其制备条件进行了优化,为进一步开展对其免疫效果和佐剂效应等研究奠定了基础。

Objective To prepare the bacterial ghost of bovine enterotoxigenic E. coli and optimize the condition for pyrolysis. Methods E. coli was transformed with lysozyme expression plasmid p HH43 and incubated at 37 ℃. When the A（600） value reached 0. 6, the temperature was increased to 42 ℃ for induction of pyrolysis and preparation of bacterial ghost. The initial A（600） for induction（0. 4, 0. 6, 0. 8, 1. 0 and 1. 2）and content of medium（2/3, 3/4, 4/5 and normal content） were optimized, and the pyrolysis rate was calculated. Results The pyrolysis rate reached the maximum of99. 93% at an initial A（600） value of 0. 6 and 3/4 of normal content of medium. Conclusion The bacterial ghost of bovine enterotoxigenic E. coli was successfully prepared, while the condition for preparation was optimized, which laid a foundation of further study on its immune and adjuvant effects.