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酿酒酵母中葡萄糖阻遏基因表达所需基因SNF1的克隆与遗传图谱绘制。

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

作者信息

Celenza J L, Carlson M

出版信息

Mol Cell Biol. 1984 Jan;4(1):49-53. doi: 10.1128/mcb.4.1.49-53.1984.

Abstract

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.

摘要

在酿酒酵母中,需要功能性的SNF1基因产物来解除对许多葡萄糖可阻遏基因的表达抑制。携带snf1突变的菌株无法在蔗糖、半乳糖、麦芽糖、蜜二糖或不可发酵碳源上生长;这些碳源的利用受葡萄糖抑制调控。snf1突变体无法利用蔗糖是由于在RNA水平上未能解除对转化酶结构基因表达的抑制。我们通过互补酿酒酵母中的snf1缺陷分离出了携带SNF1基因的重组质粒。在五个不同质粒中克隆的DNA片段有一个3.5千碱基的共同区域。用携带一段克隆DNA的整合载体转化酿酒酵母,导致质粒整合到SNF1位点。这一结果表明克隆的DNA与SNF1位点的序列同源。通过在该转化体中定位与SNF1连锁的质粒标记,我们表明SNF1基因位于第四条染色体上。然后我们将snf1定位到右臂上rna3远端5.6厘摩的位置;snf1与任何先前定位的突变都没有极其紧密的连锁关系。

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