Wards B J, Collins D M
AgResearch, Wallaceville Animal Research Centre, Upper Hutt, New Zealand.
FEMS Microbiol Lett. 1996 Nov 15;145(1):101-5. doi: 10.1111/j.1574-6968.1996.tb08563.x.
The effects of electroporation temperature, biochemical pretreatment of cells and stage of culture on electroporation efficiency for slow-growing mycobacteria were investigated. The efficiency of transformation into Mycobacterium tuberculosis, Mycobacterium bovis and Mycobacterium intracellulare increased markedly with temperature. In contrast, the efficiency of transformation into Mycobacterium smegmatis, a fast-growing species, was higher at 0 degree C and decreased with temperature. While stage of culture had little effect, a further increase in efficiency of 2-4-fold was obtained following glycine or ethionamide pretreatment. Electroporation at 37 degrees C has been chosen as a standard condition for slow-growing species as it usually resulted in a transformation efficiency several orders of magnitude higher than that obtained at 0 degree C.
研究了电穿孔温度、细胞的生化预处理以及培养阶段对生长缓慢的分枝杆菌电穿孔效率的影响。转化为结核分枝杆菌、牛分枝杆菌和胞内分枝杆菌的效率随温度显著提高。相比之下，转化为快速生长的耻垢分枝杆菌的效率在0℃时较高，并随温度降低。虽然培养阶段影响较小，但甘氨酸或乙硫异烟胺预处理后效率进一步提高了2至4倍。对于生长缓慢的菌种，已选择37℃进行电穿孔作为标准条件，因为其转化效率通常比0℃时高出几个数量级。
