Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
18
pubmed:dateCreated
1992-10-13
pubmed:databankReference
pubmed:abstractText
A model system for one-step gene disruption for an asporogenous methylotrophic yeast, Candida boidinii, is described. In this system, the 3-isopropylmalate dehydrogenase gene (C. boidinii LEU2) was selected as the target gene for disruption to derive new host strains for transformation. First, the C. boidinii LEU2 gene was cloned, and its complete nucleotide sequence was determined. Next, the LEU2 disruption vectors, which had the C. boidinii URA3 gene as the selectable marker, were constructed. Of the Ura+ transformants obtained with these plasmids, more than half showed a Leu- phenotype. Finally, the double-marker strains of C. boidinii were derived. When vectors with repeated flanking sequences of the C. boidinii URA3 gene were used for gene disruption, Leu- Ura+ transformants changed spontaneously to a Leu- Ura- phenotype ca. 100 times more frequently than they did when plasmids without the repeated sequences were used. Southern analysis showed that these events included a one-step gene disruption and a subsequent popping out of the C. boidinii URA3 sequence from the transformant chromosome.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1105072, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1195397, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1367303, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-16347563, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1875924, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1909916, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-1938943, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-2005793, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-271968, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-2760051, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-2897248, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-2985470, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3031459, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-322128, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3241624, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3241625, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3305158, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3309174, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3323803, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-347451, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-3539364, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-388356, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6280875, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6297759, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6312838, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6323436, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6336730, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-6394957, http://linkedlifedata.com/resource/pubmed/commentcorrection/1522074-765476
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Sep
pubmed:issn
0021-9193
pubmed:author
pubmed:issnType
Print
pubmed:volume
174
pubmed:geneSymbol
LEU2
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
5988-93
pubmed:dateRevised
2010-9-7
pubmed:meshHeading
pubmed:year
1992
pubmed:articleTitle
Directed mutagenesis in an asporogenous methylotrophic yeast: cloning, sequencing, and one-step gene disruption of the 3-isopropylmalate dehydrogenase gene (LEU2) of Candida boidinii to derive doubly auxotrophic marker strains.
pubmed:affiliation
Department of Agricultural Chemistry, Faculty of Agriculture, Kyoto University, Japan.
pubmed:publicationType
Journal Article, Comparative Study, Research Support, Non-U.S. Gov't