影响酿酒酵母电击转化率的条件

TRANSFORMATION OF S.CEREVISIAE BY ELECTROPORATION

以酿酒酵母为材料，用穿梭质粒ｐＹＥＳ２进行电击转化的条件实验．实验表明电击中电压大小、细胞的生长状态、处理条件和电击后细胞的培养时间以及质粒浓度等对电击转化率均有影响．取对数生长期酵母细胞，用二硫苏糖醇（ＤＴＴ）处理以疏松细胞壁，加０．７μｇ／ｍＬ质粒ＤＮＡ，５ｋＶ电压电击，转化后细胞在完全培养基中培养６０ｍｉｎ后，涂布选择培养基，转化率可达４．８×１０５个转化子／μｇ质粒ＤＮＡ，细胞的存活率为３９．２％，比完整细胞转化的效率高。

Electrotransformation was performed to transfer shuttle plasmid pYES2 into S.cerevisiae .Voltage、cell grown、incubation time and plasmid concentration affect transformation efficiency.The best conditions are the cell harvested at the exponential growth phase、5 kV、0.7 μg plasmid DNA and 1 h incubation time,transformation efficiency is up to 4.8×10 5 transformants per μg plasmid DNA,which is higher than intact cell transformation.The cell viability is about 39.2%.