期刊
FUNGAL GENETICS AND BIOLOGY
卷 29, 期 1, 页码 38-48出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1006/fgbi.1999.1180
关键词
homologous recombination; pathogenic fungi; mycology; basidiomycete
资金
- NIAID NIH HHS [R01AI42159, R01AI41937, R01AI39115] Funding Source: Medline
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 similar to 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. (C) 2000 Academic Press.
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