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酿酒酵母磷酸葡萄糖异构酶基因的分离、特性鉴定及核苷酸序列分析

The isolation, characterization and nucleotide sequence of the phosphoglucoisomerase gene of Saccharomyces cerevisiae.

作者信息

Tekamp-Olson P, Najarian R, Burke R L

机构信息

Chiron Corporation, Emeryville, CA 94608.

出版信息

Gene. 1988 Dec 15;73(1):153-61. doi: 10.1016/0378-1119(88)90321-6.

Abstract

We have isolated the gene which encodes the glycolytic enzyme phosphoglucoisomerase (PGI) from the yeast Saccharomyces cerevisiae by functional complementation of a yeast mutant deficient in PGI activity with DNA from a wild-type yeast genomic library. The cloned gene has been localized by hybridization of specific DNA fragments to total yeast poly(A)+ RNA and by complementation of the mutant phenotype with subclones. The gene is expressed as an abundant mRNA of 1.9-kb and encodes a protein of 554 amino acids with an Mr of 61310. The nucleotide sequence of the gene as well as the 5' and 3' flanking regions are presented. The predicted PGI amino acid sequence shows a high degree of homology with the sequence predicted for human and mouse neuroleukin, a putative neurotropic factor. The codon usage within the coding region is very restricted, characteristic of a highly expressed yeast gene.

摘要

我们通过用来自野生型酵母基因组文库的DNA对缺乏磷酸葡萄糖异构酶(PGI)活性的酵母突变体进行功能互补,从酿酒酵母中分离出了编码糖酵解酶磷酸葡萄糖异构酶(PGI)的基因。通过将特定DNA片段与总酵母聚腺苷酸加尾RNA杂交以及用亚克隆对突变体表型进行互补,已确定了克隆基因的位置。该基因表达为一种1.9 kb的丰富mRNA,编码一个由554个氨基酸组成、分子量为61310的蛋白质。文中给出了该基因以及5'和3'侧翼区域的核苷酸序列。预测的PGI氨基酸序列与预测的人及小鼠神经白细胞素(一种假定的神经营养因子)序列具有高度同源性。编码区内的密码子使用非常受限,这是高表达酵母基因的特征。

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