• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of iaiPermissionsJournals.ASM.orgJournalIAI ArticleJournal InfoAuthorsReviewers
Infect Immun. Mar 1992; 60(3): 876–884.
PMCID: PMC257568

Gene isolation by complementation in Candida albicans and applications to physical and genetic mapping.

Abstract

We have isolated three genes, ARG57, SER57, and LYS1, on the basis of their function in Candida albicans. A C. albicans transformation vector containing the C. albicans URA3 gene, a Candida ARS sequence, and a portion of the Saccharomyces cerevisiae 2 microns circle containing the replication origin was constructed. Clones from genomic libraries in this vector were isolated by direct complementation of the auxotrophies in strain 1006 (arg57 ser57 lys1 ura3 MPA1). Transformants typically contain two to four plasmids in a mixed tandem multimer. A scheme to resolve mixed multimers into monomers in vivo by transformation of S. cerevisiae with Candida transformant DNA selecting Ura+ transformants was devised. Monomeric plasmids were then isolated by transformation of Escherichia coli with the S. cerevisiae transformant DNA. These were retested by transformation of strain 1006 to identify the specific plasmid that complemented the auxotrophy. The chromosomal locations of the genes were determined by hybridization to C. albicans chromosomes separated on contour-clamped homogenous electric field gels. We used these locations to assess the stability of individual C. albicans chromosomes in parasexual genetic analysis. The Lys(+)-complementing clone was shown to be LYS1 by complementation of S. cerevisiae lys1 mutants. These cloned genes help to align the Candida physical and genetic maps and provide additional markers for the transformation system.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.9M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Botstein D, Falco SC, Stewart SE, Brennan M, Scherer S, Stinchcomb DT, Struhl K, Davis RW. Sterile host yeasts (SHY): a eukaryotic system of biological containment for recombinant DNA experiments. Gene. 1979 Dec;8(1):17–24. [PubMed]
  • Dutcher SK. Internuclear transfer of genetic information in kar1-1/KAR1 heterokaryons in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Mar;1(3):245–253. [PMC free article] [PubMed]
  • Gillum AM, Tsay EY, Kirsch DR. Isolation of the Candida albicans gene for orotidine-5'-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations. Mol Gen Genet. 1984;198(1):179–182. [PubMed]
  • Goshorn AK, Scherer S. Genetic analysis of prototrophic natural variants of Candida albicans. Genetics. 1989 Dec;123(4):667–673. [PMC free article] [PubMed]
  • Hilton C, Markie D, Corner B, Rikkerink E, Poulter R. Heat shock induces chromosome loss in the yeast Candida albicans. Mol Gen Genet. 1985;200(1):162–168. [PubMed]
  • Ito H, Fukuda Y, Murata K, Kimura A. Transformation of intact yeast cells treated with alkali cations. J Bacteriol. 1983 Jan;153(1):163–168. [PMC free article] [PubMed]
  • Kakar SN, Partridge RM, Magee PT. A genetic analysis of Candida albicans: isolation of a wide variety of auxotrophs and demonstration of linkage and complementation. Genetics. 1983 Jun;104(2):241–255. [PMC free article] [PubMed]
  • Kelly R, Miller SM, Kurtz MB, Kirsch DR. Directed mutagenesis in Candida albicans: one-step gene disruption to isolate ura3 mutants. Mol Cell Biol. 1987 Jan;7(1):199–208. [PMC free article] [PubMed]
  • Kurtz MB, Cortelyou MW, Kirsch DR. Integrative transformation of Candida albicans, using a cloned Candida ADE2 gene. Mol Cell Biol. 1986 Jan;6(1):142–149. [PMC free article] [PubMed]
  • Kurtz MB, Cortelyou MW, Miller SM, Lai M, Kirsch DR. Development of autonomously replicating plasmids for Candida albicans. Mol Cell Biol. 1987 Jan;7(1):209–217. [PMC free article] [PubMed]
  • Lott TJ, Page LS, Boiron P, Benson J, Reiss E. Nucleotide sequence of the Candida albicans aspartyl proteinase gene. Nucleic Acids Res. 1989 Feb 25;17(4):1779–1779. [PMC free article] [PubMed]
  • Mason MM, Lasker BA, Riggsby WS. Molecular probe for identification of medically important Candida species and Torulopsis glabrata. J Clin Microbiol. 1987 Mar;25(3):563–566. [PMC free article] [PubMed]
  • Orr-Weaver TL, Szostak JW, Rothstein RJ. Yeast transformation: a model system for the study of recombination. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354–6358. [PMC free article] [PubMed]
  • Poulter R, Jeffery K, Hubbard MJ, Shepherd MG, Sullivan PA. Parasexual genetic analysis of Candida albicans by spheroplast fusion. J Bacteriol. 1981 Jun;146(3):833–840. [PMC free article] [PubMed]
  • Poulter RT, Rikkerink EH. Genetic analysis of red, adenine-requiring mutants of Candida albicans. J Bacteriol. 1983 Dec;156(3):1066–1077. [PMC free article] [PubMed]
  • Scherer S, Davis RW. Replacement of chromosome segments with altered DNA sequences constructed in vitro. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4951–4955. [PMC free article] [PubMed]
  • Scherer S, Magee PT. Genetics of Candida albicans. Microbiol Rev. 1990 Sep;54(3):226–241. [PMC free article] [PubMed]
  • Scherer S, Stevens DA. Application of DNA typing methods to epidemiology and taxonomy of Candida species. J Clin Microbiol. 1987 Apr;25(4):675–679. [PMC free article] [PubMed]
  • Scherer S, Stevens DA. A Candida albicans dispersed, repeated gene family and its epidemiologic applications. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1452–1456. [PMC free article] [PubMed]
  • Schwartz DC, Cantor CR. Separation of yeast chromosome-sized DNAs by pulsed field gradient gel electrophoresis. Cell. 1984 May;37(1):67–75. [PubMed]
  • Shepherd MG, Poulter RT, Sullivan PA. Candida albicans: biology, genetics, and pathogenicity. Annu Rev Microbiol. 1985;39:579–614. [PubMed]
  • Smith HA, Allaudeen HS, Whitman MH, Koltin Y, Gorman JA. Isolation and characterization of a beta-tubulin gene from Candida albicans. Gene. 1988;63(1):53–63. [PubMed]
  • Suzuki T, Rogers AL, Magee PT. Inter- and intra-species crosses between Candida albicans and Candida guilliermondii. Yeast. 1986 Mar;2(1):53–58. [PubMed]
  • Tschumper G, Carbon J. High frequency excision of Ty elements during transformation of yeast. Nucleic Acids Res. 1986 Apr 11;14(7):2989–3001. [PMC free article] [PubMed]
  • Vollrath D, Davis RW. Resolution of DNA molecules greater than 5 megabases by contour-clamped homogeneous electric fields. Nucleic Acids Res. 1987 Oct 12;15(19):7865–7876. [PMC free article] [PubMed]
  • Whelan WL, Markie DM, Simpkin KG, Poulter RM. Instability of Candida albicans hybrids. J Bacteriol. 1985 Mar;161(3):1131–1136. [PMC free article] [PubMed]
  • Wickes B, Staudinger J, Magee BB, Kwon-Chung KJ, Magee PT, Scherer S. Physical and genetic mapping of Candida albicans: several genes previously assigned to chromosome 1 map to chromosome R, the rDNA-containing linkage group. Infect Immun. 1991 Jul;59(7):2480–2484. [PMC free article] [PubMed]

Articles from Infection and Immunity are provided here courtesy of American Society for Microbiology (ASM)

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links