第四条染色体上六个开放阅读框（YDL053c、YDL072c、YDL073w、YDL076c、YDL077c和YDL080c）的破坏与功能分析。
Disruption and functional analysis of six ORFs on chromosome IV: YDL053c, YDL072c, YDL073w, YDL076c, YDL077c and YDL080c.
作者信息
Blasco A, Sanz P
机构信息
Instituto de Biomedicina de Valencia (CSIC), Jaime Roig 11, 46010-Valencia, Spain.
出版信息
Yeast. 2000 Nov;16(15):1437-43. doi: 10.1002/1097-0061(200011)16:15<1437::AID-YEA633>3.0.CO;2-D.
Six open reading frames (ORFs) from chromosome IV, YDL053c, YDL072c, YDL073w, YDL076c, YDL077c and YDL080c, were disrupted using the long flanking homology technique (LFH) to replace each target locus with the KanMX4 selection marker. We have also constructed plasmids containing replacement cassettes (pYORC) and the cognate clones (pYCG) for each ORF. Disruption of five of the ORFs-YDL053c, YDL072c, YDL073w, YDL076c and YDL080c (THI3)-resulted in no distinctive phenotype with respect to temperature or nutritional requirements. However, disruption of YDL077c (also known as VAM6) exhibited an slow growth phenotype in minimal media and also in rich media containing glycerol as a carbon source. The homozygous disruptant diploid corresponding to this gene also failed to sporulate.
利用长侧翼同源技术（LFH）破坏了来自第四条染色体的六个开放阅读框（ORF），即YDL053c、YDL072c、YDL073w、YDL076c、YDL077c和YDL080c，用KanMX4选择标记替换每个目标基因座。我们还构建了包含每个ORF的替换盒（pYORC）和同源克隆（pYCG）的质粒。YDL053c、YDL072c、YDL073w、YDL076c和YDL080c（THI3）这五个ORF的破坏在温度或营养需求方面未产生明显表型。然而，YDL077c（也称为VAM6）的破坏在基本培养基以及含有甘油作为碳源的丰富培养基中表现出生长缓慢的表型。对应于该基因的纯合破坏二倍体也未能形成孢子。
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