Cloning of the Candida glabrata TRP1 and HIS3 genes, and construction of their disruptant strains by sequential integrative transformation

Gene. 1995 Nov 20;165(2):203-6. doi: 10.1016/0378-1119(95)00552-h.

Abstract

The Candida glabrata (Cg) TRP1 and HIS3 genes have been isolated by complementation of the Saccharomyces cerevisiae (Sc) trp1 and his3 mutants, respectively. Cg TRP1 encodes a polypeptide of 217 amino acids (aa), whose aa sequence is 58% identical to that of Sc TRP1. Cg HIS3 encodes a polypeptide of 210 aa, whose aa sequence is 73% identical to that of the Sc HIS3. Both Cg TRP1 and HIS3 were disrupted by sequential integrative transformation where the Sc URA3 was used as a selection marker for transformation. The resulting auxotrophic strain of his3- and trp1- was used to examine the ability of the Sc genes to complement the Cg mutations; Sc HIS3 and TRP1 complemented the Cg his3- and trp1- mutations, respectively.

MeSH terms

  • Aldose-Ketose Isomerases*
  • Amino Acid Sequence
  • Base Sequence
  • Candida / genetics*
  • Cloning, Molecular
  • Fungal Proteins / genetics*
  • Genes, Fungal / genetics*
  • Genetic Complementation Test
  • Hydro-Lyases / genetics*
  • Molecular Sequence Data
  • Mutagenesis*
  • Restriction Mapping
  • Saccharomyces cerevisiae / genetics
  • Saccharomyces cerevisiae Proteins*
  • Sequence Analysis, DNA
  • Sequence Homology, Amino Acid
  • Transformation, Genetic

Substances

  • Fungal Proteins
  • Saccharomyces cerevisiae Proteins
  • Hydro-Lyases
  • imidazoleglycerolphosphate dehydratase
  • Aldose-Ketose Isomerases
  • TRP1 protein, S cerevisiae

Associated data

  • GENBANK/U31470
  • GENBANK/U31471