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Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans.

Publication ,  Journal Article
Davidson, RC; Cruz, MC; Sia, RA; Allen, B; Alspaugh, JA; Heitman, J
Published in: Fungal Genet Biol
February 2000

Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans. Fungal Genetics and Biology 29, 38-48. Cryptococcus neoformans is an opportunistic fungal pathogen with a defined sexual cycle and well-developed genetic and molecular approaches. Two different transformation systems have been developed, and a number of genes have been disrupted by homologous recombination. However, the frequency of homologous recombination achieved by these approaches has differed dramatically between strains of the A and D serotypes. Transformation by electroporation in serotype D strains results in homologous recombination at frequencies of 1/1000 to 1/100,000, whereas transformation by the biolistic method has resulted in gene disruption at frequencies between 2 and 50% in serotype A strains. We find that gene disruption by homologous recombination can be achieved in the congenic serotype D strain series by biolistic transformation with frequencies of approximately 1 to 4%. By this approach, we have readily disrupted the genes encoding a MAPK homolog (CPK1), the calcineurin A catalytic subunit (CNA1), and a G protein alpha subunit (GPA1). By physical and genetic methods, we show that these mutations result from targeted recombination events without ectopic integrations. Because genetic approaches can be applied in the congenic serotype D strains, our observations represent a significant advance in molecular approaches to understand the physiology and virulence of this important human pathogen.

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Published In

Fungal Genet Biol

DOI

ISSN

1087-1845

Publication Date

February 2000

Volume

29

Issue

1

Start / End Page

38 / 48

Location

United States

Related Subject Headings

  • Transformation, Genetic
  • Tacrolimus Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Recombination, Genetic
  • Plasmids
  • Phosphoprotein Phosphatases
  • Mitogen-Activated Protein Kinases
  • Microbiology
  • Immunophilins
  • Heterotrimeric GTP-Binding Proteins
 

Citation

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Davidson, R. C., Cruz, M. C., Sia, R. A., Allen, B., Alspaugh, J. A., & Heitman, J. (2000). Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans. Fungal Genet Biol, 29(1), 38–48. https://doi.org/10.1006/fgbi.1999.1180
Davidson, R. C., M. C. Cruz, R. A. Sia, B. Allen, J. A. Alspaugh, and J. Heitman. “Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans.Fungal Genet Biol 29, no. 1 (February 2000): 38–48. https://doi.org/10.1006/fgbi.1999.1180.
Davidson RC, Cruz MC, Sia RA, Allen B, Alspaugh JA, Heitman J. Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans. Fungal Genet Biol. 2000 Feb;29(1):38–48.
Davidson, R. C., et al. “Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans.Fungal Genet Biol, vol. 29, no. 1, Feb. 2000, pp. 38–48. Pubmed, doi:10.1006/fgbi.1999.1180.
Davidson RC, Cruz MC, Sia RA, Allen B, Alspaugh JA, Heitman J. Gene disruption by biolistic transformation in serotype D strains of Cryptococcus neoformans. Fungal Genet Biol. 2000 Feb;29(1):38–48.
Journal cover image

Published In

Fungal Genet Biol

DOI

ISSN

1087-1845

Publication Date

February 2000

Volume

29

Issue

1

Start / End Page

38 / 48

Location

United States

Related Subject Headings

  • Transformation, Genetic
  • Tacrolimus Binding Proteins
  • Saccharomyces cerevisiae Proteins
  • Recombination, Genetic
  • Plasmids
  • Phosphoprotein Phosphatases
  • Mitogen-Activated Protein Kinases
  • Microbiology
  • Immunophilins
  • Heterotrimeric GTP-Binding Proteins