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Applied Microbiology and Biotechnology

, Volume 44, Issue 3–4, pp 444–450 | Cite as

Genetic transformation of the pathogenic fungus Wangiella dermatitidis

  • M. Peng
  • C. R. CooperJr.
  • P. J. Szaniszlo
Applied Genetics and Regulation Original Paper

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/μg pAN7-1 on medium containing 100 μg HmB/ml. Strains of W. dermatitidis used in this study have innate sensitivity to HmB at a critical inhibitory concentration of 20–40 μg/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/μg vector DNA. Southern blotting analysis and polymerase chain reaction detection of DNA from putative transformants confirmed transformation.

Keywords

Polymerase Chain Reaction Southern Blotting Pathogenic Fungus Hygromycin Southern Blotting Analysis 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • M. Peng
    • 1
  • C. R. CooperJr.
    • 2
  • P. J. Szaniszlo
    • 1
  1. 1.Department of MicrobiologyUniversity of Texas at AustinAustinUSA
  2. 2.Department of PathologyUniversity of Texas Medical BranchGalvestonUSA

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