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Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA.

Publication ,  Journal Article
Toffaletti, DL; Rude, TH; Johnston, SA; Durack, DT; Perfect, JR
Published in: J Bacteriol
March 1993

A transformation scheme for Cryptococcus neoformans to yield high-frequency, integrative events was developed. Adenine auxotrophs from a clinical isolate of C. neoformans serotype A were complemented by the cryptococcal phosphoribosylaminoimidazole carboxylase gene (ade2) with a biolistic DNA delivery system. Comparison of two DNA delivery systems (electroporation versus a biolistic system) showed notable differences. The biolistic system did not require linear vectors and transformed each auxotrophic strain at similar frequencies. Examination of randomly selected transformants by biolistics showed that 15 to 40% were stable, depending on the recipient auxotroph, with integrative events identified in all stable transformants by DNA analysis. Although the ade2 cDNA copy transformed at a low frequency, DNA analysis found homologous recombination in each of these transformants. DNA analysis of stable transformants receiving genomic ade2 revealed ectopic integration in a majority of cases, but approximately a quarter of the transformants showed homologous recombination with vector integration or gene replacement. This system has the potential for targeted gene disruption, and its efficiency will also allow for screening of DNA libraries within C. neoformans. Further molecular strategies to study the pathobiology of this pathogenic yeast are now possible with this transformation system.

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Published In

J Bacteriol

DOI

ISSN

0021-9193

Publication Date

March 1993

Volume

175

Issue

5

Start / End Page

1405 / 1411

Location

United States

Related Subject Headings

  • Transformation, Bacterial
  • Transfection
  • Restriction Mapping
  • Mitosis
  • Microbiology
  • Genetic Complementation Test
  • Cryptococcus neoformans
  • Carboxy-Lyases
  • Blotting, Southern
  • 11 Medical and Health Sciences
 

Citation

APA
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Toffaletti, D. L., Rude, T. H., Johnston, S. A., Durack, D. T., & Perfect, J. R. (1993). Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA. J Bacteriol, 175(5), 1405–1411. https://doi.org/10.1128/jb.175.5.1405-1411.1993
Toffaletti, D. L., T. H. Rude, S. A. Johnston, D. T. Durack, and J. R. Perfect. “Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA.J Bacteriol 175, no. 5 (March 1993): 1405–11. https://doi.org/10.1128/jb.175.5.1405-1411.1993.
Toffaletti DL, Rude TH, Johnston SA, Durack DT, Perfect JR. Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA. J Bacteriol. 1993 Mar;175(5):1405–11.
Toffaletti, D. L., et al. “Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA.J Bacteriol, vol. 175, no. 5, Mar. 1993, pp. 1405–11. Pubmed, doi:10.1128/jb.175.5.1405-1411.1993.
Toffaletti DL, Rude TH, Johnston SA, Durack DT, Perfect JR. Gene transfer in Cryptococcus neoformans by use of biolistic delivery of DNA. J Bacteriol. 1993 Mar;175(5):1405–1411.

Published In

J Bacteriol

DOI

ISSN

0021-9193

Publication Date

March 1993

Volume

175

Issue

5

Start / End Page

1405 / 1411

Location

United States

Related Subject Headings

  • Transformation, Bacterial
  • Transfection
  • Restriction Mapping
  • Mitosis
  • Microbiology
  • Genetic Complementation Test
  • Cryptococcus neoformans
  • Carboxy-Lyases
  • Blotting, Southern
  • 11 Medical and Health Sciences