Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
20
pubmed:dateCreated
1994-11-23
pubmed:abstractText
For characterizing in vivo functions of a mammalian protein, it is informative to obtain conditional mutations and apply them to the mouse genetic system. However, the isolation of conditional mutations has been quite difficult in cultured cells. We report here that functional expression of a heterologous mammalian gene in the yeast Saccharomyces cerevisiae provides a system for isolating mutated genes. We found that the cloned mouse TOP2 alpha cDNA, which encodes mouse DNA topoisomerase II (topo II) alpha, could rescue the lethal phenotype caused by yeast top2 null mutation. In order to generate and select temperature-sensitive mouse topo II alpha, an expression plasmid was mutagenized in vitro and was transformed, using the plasmid shuffling method, into the yeast strain, in which the endogenous TOP2 gene had been disrupted. We observed that one of such clone of yeast cells harboring a mutagenized mouse TOP2 alpha showed temperature-sensitive growth. Enzymatic assays and sequencing analysis revealed that this phenotype was caused by the thermosensitive nature of the mutant mouse protein, which has isoleucine at amino acid 961 instead of threonine. Therefore we have isolated the first conditional mutation in the mouse TOP2 alpha.
pubmed:commentsCorrections
http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-1331984, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-1333583, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-15957215, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-1655564, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2005795, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2107073, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2197185, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2544296, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2549254, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2556712, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2557897, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2659436, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-271968, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2824504, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2840207, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2845399, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2902925, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2985283, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2985625, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2987966, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2992360, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-2999980, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-3016287, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-3023070, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-3040264, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-3289117, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-3319781, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-375221, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6090122, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6276569, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6323017, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6326134, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6336730, http://linkedlifedata.com/resource/pubmed/commentcorrection/7937150-6394957
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0305-1048
pubmed:author
pubmed:issnType
Print
pubmed:day
11
pubmed:volume
22
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
4229-33
pubmed:dateRevised
2010-11-18
pubmed:meshHeading
pubmed:year
1994
pubmed:articleTitle
Mutant isolation of mouse DNA topoisomerase II alpha in yeast.
pubmed:affiliation
Institute of Medical Science, University of Tokyo, Japan.
pubmed:publicationType
Journal Article, Research Support, Non-U.S. Gov't