电穿孔方法是一种高效导入外源DNA的技术。通过电转化法对酵母YPH 500进行外源DNA转化,研究了电转化中各因素对酵母转化率的影响。结果表明,电场强度、外源DNA浓度、感受态细胞状态对转化率有明显影响。采用均匀设计试验,利用逐步回归方法对试验结果进行分析,结果表明,当电场强度在1750V,酵母OD600nm在1.3,外源DNA浓度为7.5 mg/L时,酵母转化率较高,约为24×102转化子数/μg DNA。
Electroporation is a highly efficient technique introducing extraneous DNA. The method was used to transform Saccharomyces cerevisiae YPH 500. The effects of different factors on the electrotransformation were studied. The results showed that the electric intensity, the concentration of extraneous DNA, and the status of competence significantly affected on transformation efficiency. Homogeneous design experiment was used to analyze by stepwise regression method. The results showed that the transformation ratio of Saccharomyces cerevisiae YPH 500 was 24 ×102transformations/μg DNA under the electric intensity of 1 750 V, OD600 nmof 1.3 and the concentration of extraneous DNA of 7.5 mg/L.