乳酸乳球菌电转化条件的研究

The factors affected transformation efficiency of Lactococcus lactis by electroporation

以质粒pTRKH2为材料,对乳酸乳球菌MG1614进行电转化条件的研究.实验结果表明,在电转化过程中,电场强度、电阻大小、细胞生长状态和电击后菌株复苏时间均对转化效率有明显影响,得到了乳酸乳球菌的最适电转化条件为:对数生长中期的细胞,经1%甘氨酸处理,在10kVcm电场强度、400Ω电阻下电转化,转化后细胞在SGM17培养液中培养2h后涂布选择性培养基,转化率(转化子个数与DNA质量之比)可达1.2×103个μg,比优化前的转化效率提高近100倍.

Electrotransformation factors were performed to transfer plasmid pTRKH_2 into Lactococcus lactis MG1614. Cell growth phase, electric field strength, resistance and incubation time all affect transformation efficiency. The optimum factors are the cell harvested at the middle exponential growth phase, electroporation at 10.0kV/cm and 400Ω, 2h cell incubation time. The transformation efficiency goes up to 1.2×10 3 transformants per μg DNA with those optimum factors, which is higher than optimiz those reported before.