López M C, Sánchez M, Fermiñán E, Domínguez A
Departamento de Microbiología y Genética, Universidad de Salamanca/CSIC, Spain.
Yeast. 1998 Sep 30;14(13):1199-208. doi: 10.1002/(SICI)1097-0061(19980930)14:13<1199::AID-YEA309>3.0.CO;2-V.
We report here the construction of six deletion mutants and the analysis of their basic phenotype. Deletion cassettes containing the KanMX4 marker module and long flanking regions homologous to the target locus were constructed for each of the six open reading-frames (ORFs YDL088c, YDL087c, YDL086w, YDL085w, YDL084w and YDL082w) located on chromosome IV. Sporulation and tetrad analysis of heterozygous deletant strains revealed that, in the FY1679 genetic background, ORFs YDL088c, YDL087c and YDL084w are essential genes for vegetative growth whereas YDL086w, YDL085w and YDL082w are non-essential. ydl088c delta and ydl084w delta haploid strains are viable in the CEN. PK2 genetic background although ydl084w delta grows at a slower rate than the wild type. Complementation tests by corresponding cognate genes confirmed that gene inactivation was responsible for these growth defects.
我们在此报告六个缺失突变体的构建及其基本表型分析。针对位于第四条染色体上的六个开放阅读框（ORF YDL088c、YDL087c、YDL086w、YDL085w、YDL084w和YDL082w），构建了包含KanMX4标记模块以及与目标位点同源的长侧翼区域的缺失盒。杂合缺失菌株的孢子形成和四分体分析表明，在FY1679遗传背景下，ORF YDL088c、YDL087c和YDL084w是营养生长的必需基因，而YDL086w、YDL085w和YDL082w是非必需基因。ydl088c delta和ydl084w delta单倍体菌株在CEN.PK2遗传背景下是可行的，尽管ydl084w delta的生长速度比野生型慢。通过相应同源基因进行的互补试验证实，基因失活是这些生长缺陷的原因。
