Isolation of the Candida albicans gene for orotidine-5′-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations
- 1 December 1984
- journal article
- research article
- Published by Springer Science and Business Media LLC in Molecular Genetics and Genomics
- Vol. 198 (1), 179-182
- https://doi.org/10.1007/bf00328721
Abstract
A gene bank of Sau3A partially digested Candida albicans DNA in vector YEp13 was used to complement a ura3 mutation (orotidine-5′-phosphate decarboxylase, OMPdecase) in S. cerevisiae. Two plasmids which complemented ura3 and showed clear linkage of Ura+ and plasmid markers were selected for further study. Both plasmids also complemented the corresponding OMPdecase mutation (pyrF) in E. coli. Restriction mapping and subcloning studies localized the OMPdecase complementing activity to a region common to both plasmids. Probes prepared from this common region hybridized specifically to C. albicans DNA and not to E. coli or S. cerevisiae DNA. Southern blot analysis also showed that the restriction map of the ura3 complementing region of one plasmid was colinear with C. albicans genomic DNA. Expression of the OMPdecase complementing gene in E. coli and S. cerevisiae was not dependent upon orientation relative to vector sequences, suggesting that promotion could be occurring within the C. albicans fragment. Expression was sufficient to allow complementation in S. cerevisiae with integrating as well as high copy number vectors.Keywords
This publication has 26 references indexed in Scilit:
- Hybridization of Candida albicans through fusion of protoplastsArchiv für Mikrobiologie, 1981
- Analysis of the Structure and Function of the Kanamycin-resistance Transposon Tn903Cold Spring Harbor Symposia on Quantitative Biology, 1981
- Heterozygosity and segregation in Candida albicansMolecular Genetics and Genomics, 1980
- Transformation in yeast: Development of a hybrid cloning vector and isolation of the can1 geneGene, 1979
- Sterile host yeasts (SHY): A eukaryotic system of biological containment for recombinant DNA experimentsGene, 1979
- Insertion of a genetic marker into the ribosomal DNA of yeastPlasmid, 1979
- Construction and characterization of new cloning vehicles III. Derivatives of plasmid pBR322 carrying unique Eco RI sites for selection of Eco RI generated recombinant DNA moleculesGene, 1978
- Isolation and analysis of recombinant DNA molecules containing yeast DNAGene, 1978
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- Detection of specific sequences among DNA fragments separated by gel electrophoresisJournal of Molecular Biology, 1975