Genetic transformation of the pathogenic fungus Wangiella dermatitidis

Applied Microbiology and Biotechnology
M PengP J Szaniszlo

Abstract

Genetic transformation of Wangiella dermatitidis was studied using three plasmid vectors (pAN7-1, pWU44, and pKK5) and both electroporation and polyethyleneglycol-mediated methods. pAN7-1 contains the E. coli hygromycin B (HmB) phosphotransferase (hph) gene. Expression of the hph gene confers resistance to antibiotic HmB. Selection for resistance, indicative of transformation, resulted in 10 203 HmB-resistant colonies/micrograms pAN7-1 on medium containing 100 micrograms HmB/ml. Strains of W. dermatitidis used in this study have innate sensitivity to HmB at a critical inhibitory concentration of 20-40 micrograms/ml. Vectors pWU44 and pKK5 contain a URA5 gene from Podospora anserina. A ura5 auxotroph of W. dermatitidis was transformed to prototrophy with pWU44 or pKK5 by complementation. Transformation frequencies for these two plasmids were between 17-50 transformants/micrograms vector DNA. Southern blotting analysis and polymerase chain reaction detection of DNA from putative transformants confirmed transformation.

References

Aug 1, 1978·Journal of Bacteriology·R L Roberts, P J Szaniszlo
Jan 1, 1992·Biotechnology·D B Finkelstein

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Citations

Apr 22, 2003·Infection and Immunity·P T MageeDana Davis
Nov 28, 2002·International Journal of Medical Microbiology : IJMM·Paul J Szaniszlo
Nov 10, 2001·Annual Review of Phytopathology·Joan M HensonAlan W Day

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