外源DNA导入酿酒酵母(Saccharomyces cerevisiae)2.558电转化条件的研究

Research on Conditions of Electric Conversion of Transforming Extraneous DNA into Saccharomyces cerevisiae 2.558

[目的]探讨外源DNA导入酿酒酵母(Saccharomy cescerevisiae)2.558的电转化条件,初步确立提高其转化效率的方法。[方法]通过调整电转化的参数,将已构建好的转化载体转化至酿酒酵母2.558的感受态细胞中,对影响电转化的主要因素进行研究。[结果]采用对数生长中期菌体制备的感受态细胞,在质粒DNA加入量为1μg、电场强度为1.8 kV/cm、电阻为225Ω、用复苏缓冲液Ⅱ进行复苏培养的条件下,电转化效率达到最大值。[结论]该研究确定了酿酒酵母(Saccharomy cescerevisiae)2.558的最优电转化条件,为其遗传改造奠定了良好的技术基础。

[ Objective ] The aim was to discuss the conditions of electric conversion of transforming extraneous DNA into S. cerevisiae 2. 558 and initially establish an efficient method for improving transformation efficiency. [ Methods] Through adjusting the parameters of electric conversion, the constructed transformation vectors were constructed into competent cells of S. cerevisiae 2. 558, then, the main factors which af- fected electric conversion were studied. [ Result ] Using competent cells which prepared from thalli during the metaphase of logarithmic growth, under the conditions of 1μg plasmid DNA concentration, 1.8 kV/cm electric field strength, 225 Ω resistance and resuscitative buffer Ⅱ, the electric conversion efficiency was best. [ Conclusion] The study confirmed the best electric conversion conditions of S. cerevisiae 2. 558, which provided better technological base for its genetic alteration.