Development of transformation system for Trichophyton rubrum by electroporation of germinated conidia

• DOI: 10.1007/s00294-009-0264-8
• Publication Date: 2009-07-22T11:43:49Z
• Authors: Anita Dobrowolska; Pawel Staczek
• Keywords: Blotting, Southern; Phosphotransferases (Alcohol Group Acceptor); 0303 health sciences; 03 medical and health sciences; Electroporation; Transformation, Genetic; Base Sequence; Trichophyton; Spores, Fungal; DNA Primers

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.