Dobrowolska Anita, Staczek Pawel
Department of Genetics of Microorganisms, University of Lodz, Banacha 12/16, 90-237, Lodz, Poland.
Curr Genet. 2009 Oct;55(5):537-42. doi: 10.1007/s00294-009-0264-8. Epub 2009 Jul 23.
Dermatophytes are the fungi that can cause infections of skin, hair, and nails due to their ability to utilize keratin. The genetic transformation systems of dermatophytes were successfully applied to Trichophyton mentagrophytes and Microsporum canis. Here we describe the procedure for genetic transformation of Trichophyton rubrum by electroporation of their germinated conidia. A linearized transformation vector (pCHSH75-Pch/GFP/TtrpC) containing bacterial hygromycin B phosphotransferase gene (hph) and green fluorescent protein gene (egfp) was introduced into the germinated conidia of T. rubrum by electroporation. PCR reaction analysis showed that egfp gene was integrated randomly and Southern blotting analysis demonstrated a single integration of hph gene into the chromosomal DNA of randomly selected transformant. In this work we report the efficient transformation and selection of the stable T. rubrum transformants.
皮肤癣菌是一类能够利用角蛋白从而引起皮肤、毛发和指甲感染的真菌。皮肤癣菌的遗传转化系统已成功应用于须癣毛癣菌和犬小孢子菌。在此，我们描述了通过对红色毛癣菌已萌发的分生孢子进行电穿孔来实现其遗传转化的过程。一个含有细菌潮霉素B磷酸转移酶基因（hph）和绿色荧光蛋白基因（egfp）的线性化转化载体（pCHSH75-Pch/GFP/TtrpC）通过电穿孔被导入到红色毛癣菌已萌发的分生孢子中。PCR反应分析表明egfp基因是随机整合的，Southern印迹分析则证明hph基因单拷贝整合到了随机挑选的转化体的染色体DNA中。在这项工作中，我们报道了红色毛癣菌稳定转化体的高效转化和筛选。
