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Cloning and genetic mapping of SNF1 a gene required for expression of glucose-repressible genes in Saccharomyces cerevisiae.

机译:SNF1的克隆和基因作图SNF1是在酿酒酵母中表达葡萄糖可抑制基因所需的基因。

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摘要

A functional SNF1 gene product is required to derepress expression of many glucose-repressible genes in Saccharomyces cerevisiae. Strains carrying a snf1 mutation are unable to grow on sucrose, galactose, maltose, melibiose, or nonfermentable carbon sources; utilization of these carbon sources is regulated by glucose repression. The inability of snf1 mutants to utilize sucrose results from failure to derepress expression of the structural gene for invertase at the RNA level. We isolated recombinant plasmids carrying the SNF1 gene by complementation of the snf1 defect in S. cerevisiae. A 3.5-kilobase region is common to the DNA segments cloned in five different plasmids. Transformation of S. cerevisiae with an integrating vector carrying a segment of the cloned DNA resulted in integration of the plasmid at the SNF1 locus. This result indicates that the cloned DNA is homologous to sequences at the SNF1 locus. By mapping a plasmid marker linked to SNF1 in this transformant, we showed that the SNF1 gene is located on chromosome IV. We then mapped snf1 to a position 5.6 centimorgans distal to rna3 on the right arm; snf1 is not extremely closely linked to any previously mapped mutation.
机译:需要功能性SNF1基因产物来抑制酿酒酵母中许多葡萄糖可抑制基因的表达。携带snf1突变的菌株无法在蔗糖,半乳糖,麦芽糖,蜜三糖或不可发酵的碳源上生长;这些碳源的利用受到葡萄糖抑制的调节。 snf1突变体无法利用蔗糖的原因是无法在RNA水平上抑制转化酶的结构基因表达。我们通过酿酒酵母中snf1缺陷的互补分离了携带SNF1基因的重组质粒。在五个不同质粒中克隆的DNA片段共有3.5个碱基的区域。用携带克隆的DNA片段的整合载体转化酿酒酵母,导致质粒在SNF1基因座整合。该结果表明,克隆的DNA与SNF1基因座的序列同源。通过在此转化体中定位与SNF1连接的质粒标记,我们表明SNF1基因位于第IV染色体上。然后,我们将snf1映射到右臂上rna3远端5.6厘摩的位置。 snf1与以前映射的任何突变都没有非常紧密的联系。

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