De Backer M D, Maes D, Vandoninck S, Logghe M, Contreras R, Luyten W H
Department of Advanced Biotechnologies, Janssen Research Foundation, Turnhoutseweg 30, B2340 Beerse, Belgium.
Yeast. 1999 Nov;15(15):1609-18. doi: 10.1002/(sici)1097-0061(199911)15:15<1609::aid-yea485>3.3.co;2-p.
In contrast to a variety of other yeasts, Candida albicans has proved difficult to transform with high efficiency. Lithium acetate transformation is fast and simple but provides a very low efficiency of DNA transfer (50-100 transformants/microg DNA), while spheroplast transformation, although more efficient ( approximately 300 transformants/microg integrative DNA and 10(3)-10(4) transformants/microg replicative DNA), is complicated and time-consuming. In this study we applied various yeast transformation techniques to C. albicans and selected an electroporation procedure for further optimization. Transformation efficiencies of up to 300 transformants/microg were obtained for an integrative plasmid and up to 4500 transformants/microg for a CARS-carrying plasmid. This reasonably high transformation efficiency, combined with the ease and speed of electroporation in comparison to alternative techniques, make it the preferred method for transformation of C. albicans.
与其他多种酵母不同,白色念珠菌已被证明难以高效转化。醋酸锂转化快速简便,但DNA转移效率非常低(每微克DNA产生50 - 100个转化子),而原生质球转化虽然效率更高(每微克整合型DNA约产生300个转化子,每微克复制型DNA产生10³ - 10⁴个转化子),但复杂且耗时。在本研究中,我们将各种酵母转化技术应用于白色念珠菌,并选择了电穿孔程序进行进一步优化。对于整合质粒,每微克可获得高达300个转化子的转化效率,对于携带CARS的质粒,每微克可获得高达4500个转化子的转化效率。与其他替代技术相比,这种相当高的转化效率,再加上电穿孔的简便性和速度,使其成为白色念珠菌转化的首选方法。