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首页> 外文期刊>Journal of bacteriology >Gene-enzyme relationships in the proline biosynthetic pathway of Saccharomyces cerevisiae.
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Gene-enzyme relationships in the proline biosynthetic pathway of Saccharomyces cerevisiae.

机译：酿酒酵母脯氨酸生物合成途径中的基因-酶关系。

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摘要

The PRO1, PRO2, and PRO3 genes were isolated by functional complementation of pro1, pro2, and pro3 (proline-requiring) strains of Saccharomyces cerevisiae. Independent clones with overlapping inserts were isolated from S. cerevisiae genomic libraries in YEp24 (2 microns) and YCp50 (CEN) plasmids. The identity of each gene was determined by gene disruption, and Southern hybridization and genetic analyses confirmed that the bona fide genes had been cloned. Plasmids containing each gene were introduced into known bacterial proline auxotrophs, and the ability to restore proline prototrophy was assessed. Interspecies complementation demonstrated that the S. cerevisiae PRO1 gene encoded gamma-glutamyl kinase, PRO2 encoded gamma-glutamyl phosphate reductase, and PRO3 encoded delta 1-pyrroline-5-carboxylate reductase. The presence of the PRO3 gene on a high-copy-number plasmid in S. cerevisiae caused a 20-fold overproduction of delta 1-pyrroline-5-carboxylate reductase. The PRO2 gene mapped on chromosome XV tightly linked to cdc66, and the PRO3 gene was located on the right arm of chromosome V between HIS1 and the centromere.
机译：通过酿酒酵母的pro1，pro2和pro3（需要脯氨酸）菌株的功能互补来分离PRO1，PRO2和PRO3基因。在YEp24（2微米）和YCp50（CEN）质粒中，从啤酒酵母基因组文库中分离出具有重叠插入片段的独立克隆。每个基因的身份通过基因破坏来确定，Southern杂交和遗传分析证实了真正的基因已经被克隆。将包含每个基因的质粒引入已知的细菌脯氨酸营养缺陷型，并评估还原脯氨酸原养能力。种间互补表明酿酒酵母PRO1基因编码γ-谷氨酰激酶，PRO2编码γ-谷氨酰磷酸还原酶，而PRO3编码δ1-吡咯啉-5-羧酸还原酶。酿酒酵母中高拷贝数质粒上PRO3基因的存在导致δ1-吡咯啉-5-羧酸还原酶的过量生产20倍。定位在XV染色体上的PRO2基因与cdc66紧密相连，而PRO3基因位于HIS1与着丝粒之间的V染色体的右臂上。

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