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来自酿酒酵母的GAL80基因的分子克隆及gal80缺失的表征

Molecular cloning of the GAL80 gene from Saccharomyces cerevisiae and characterization of a gal80 deletion.

作者信息

Yocum R R, Johnston M

出版信息

Gene. 1984 Dec;32(1-2):75-82. doi: 10.1016/0378-1119(84)90034-9.

Abstract

An integrated GAL1-lacZ fusion provided a useful phenotypic marker for the gal80- regulatory mutation in Saccharomyces cerevisiae. On minimal glucose plates containing a beta-galactosidase indicator, a GAL80 strain containing the fusion gave white colonies, whereas a gal80- strain gave blue colonies. This color difference was used to isolate the GAL80 gene from a plasmid bank by complementation of the gal80- mutant. The putative GAL80 gene was located on a 2.6-kb HindIII-SalI fragment and has been subcloned into an integrating vector. Genetic analysis showed that the clone integrated at the GAL80 locus. A deletion that covered the entire GAL80 region was constructed in vitro and transplaced into the yeast genome to give an isogenic pair of GAL80 and gal80 deletion strains. Glucose repression of a GAL1-lacZ fusion was normal in the gal80 deletion strain, implying that the GAL80 gene product is not involved in glucose repression.

摘要

一个整合的GAL1-lacZ融合体为酿酒酵母中的gal80调控突变提供了一个有用的表型标记。在含有β-半乳糖苷酶指示剂的基本葡萄糖平板上,含有该融合体的GAL80菌株产生白色菌落,而gal80-菌株产生蓝色菌落。这种颜色差异被用于通过gal80-突变体的互补作用从质粒文库中分离GAL80基因。推定的GAL80基因位于一个2.6 kb的HindIII-SalI片段上,并已亚克隆到一个整合载体中。遗传分析表明,该克隆整合在GAL80位点。在体外构建了一个覆盖整个GAL80区域的缺失,并将其转座到酵母基因组中,以产生一对同基因的GAL80和gal80缺失菌株。在gal80缺失菌株中,GAL1-lacZ融合体的葡萄糖阻遏作用正常,这意味着GAL80基因产物不参与葡萄糖阻遏。

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