Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation

A. M. Celis, A. M. Vos, S. Triana, C. A. Medina, N. Escobar, S. Restrepo, H. A.B. Wösten, H. de Cock

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7 days at 19 °C and 24 °C, respectively. Transformation efficiency was 0.75–1.5% for M. furfur and 0.6–7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.

Original languageEnglish (US)
Pages (from-to)1-6
Number of pages6
JournalJournal of Microbiological Methods
Volume134
DOIs
StatePublished - Mar 1 2017
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2017

Keywords

  • Agrobacterium
  • GFP
  • Hygromycin
  • Malassezia spp.
  • T-DNA
  • Transformation

Fingerprint

Dive into the research topics of 'Highly efficient transformation system for Malassezia furfur and Malassezia pachydermatis using Agrobacterium tumefaciens-mediated transformation'. Together they form a unique fingerprint.

Cite this