Legrain P, Chapon C, Schwob E, Martin R, Rosbash M, Dujon B
Department de Biologie Moléculaire, Institut Pasteur, Paris, France.
Mol Gen Genet. 1991 Feb;225(2):199-202. doi: 10.1007/BF00269848.
In the yeast Saccharomyces cerevisiae, some thermosensitive (ts) mutants have been shown to be impaired in pre-mRNA splicing (prp mutants). From a yeast genomic library, we have isolated plasmids that complement prp6 or prp9 ts mutations. These plasmids also complement the ts growth defect of additional independent mutants identified as new prp6 and prp9 ts alleles, indicating that the cloned DNAs encode PRP6 and PRP9 genes, respectively. Here, we describe the restriction maps of these loci which are localized on chromosome II and IV, respectively. The limits of open reading frames (ORFs) within the cloned inserts have been determined using a linker insertion strategy combined with the ts complementation assay. Double-strand DNA sequencing was also performed directly on the yeast expression vector from the inserted linkers. Gene disruption experiments demonstrate that both genes are essential for viability.
在酿酒酵母中，一些温度敏感（ts）突变体已被证明在前体mRNA剪接过程中存在缺陷（prp突变体）。我们从酵母基因组文库中分离出了能够互补prp6或prp9 ts突变的质粒。这些质粒还能互补被鉴定为新的prp6和prp9 ts等位基因的其他独立突变体的ts生长缺陷，这表明克隆的DNA分别编码PRP6和PRP9基因。在此，我们描述了这些分别定位在第二条和第四条染色体上的基因座的限制性图谱。利用接头插入策略结合ts互补分析，确定了克隆插入片段内的开放阅读框（ORF）界限。还直接对插入接头后的酵母表达载体进行了双链DNA测序。基因破坏实验表明这两个基因对细胞活力都是必需的。
