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

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