Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
3
pubmed:dateCreated
1993-9-16
pubmed:abstractText
Genetic manipulation of Candida albicans is constrained by its diploid genome and asexual life cycle. Recessive mutations are not expressed when heterozygous and undesired mutations introduced in the course of random mutagenesis cannot be removed by genetic back-crossing. To circumvent these problems, we developed a genotypic screen that permitted identification of a heterozygous recessive mutation at the URA3 locus. The mutation was introduced by targeted mutagenesis, homologous integration of transforming DNA, to avoid introduction of extraneous mutations. The ura3 mutation was rendered homozygous by a second round of transformation resulting in a Ura- strain otherwise isogenic with the parental clinical isolate. Subsequent mutation of the Ura- strain was achieved by targeted mutagenesis using the URA3 gene as a selectable marker. URA3 selection was used repeatedly for the sequential introduction of mutations by flanking the URA3 gene with direct repeats of the Salmonella typhimurium hisG gene. Spontaneous intrachromosomal recombination between the flanking repeats excised the URA3 gene restoring a Ura- phenotype. These Ura- segregants were selected on 5-fluoroorotic acid-containing medium and used in the next round of mutagenesis. To permit the physical mapping of disrupted genes, the 18-bp recognition sequence of the endonuclease I-SceI was incorporated into the hisG repeats. Site-specific cleavage of the chromosome with I-SceI revealed the position of the integrated sequences.
pubmed:grant
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-1569935, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-1569957, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-1651496, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2011508, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2019440, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2201296, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2407612, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2407719, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2556711, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2574635, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2671651, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2693597, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-2842757, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3023819, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3031460, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3174435, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3282237, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3305158, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-3431451, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-6394957, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-6394964, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-6757662, http://linkedlifedata.com/resource/pubmed/commentcorrection/8349105-6780534
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jul
pubmed:issn
0016-6731
pubmed:author
pubmed:issnType
Print
pubmed:volume
134
pubmed:geneSymbol
ECE1, URA3, hisG
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
717-28
pubmed:dateRevised
2009-11-18
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Isogenic strain construction and gene mapping in Candida albicans.
pubmed:affiliation
Department of Microbiology and Molecular Genetics, California College of Medicine, University of California, Irvine 92717.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.