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首页> 外文期刊>Molecular and Cellular Biology >Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli.
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Expression of the gene for ornithine decarboxylase of Saccharomyces cerevisiae in Escherichia coli.

机译：啤酒酵母的鸟氨酸脱羧酶基因在大肠杆菌中的表达。

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摘要

Diploid cells of Saccharomyces cerevisiae homozygous for the spe1A mutation, which eliminates ornithine decarboxylase activity, were found to sporulate at a greatly reduced frequency in the absence of polyamines. Plasmids which complement the spe1A mutation were isolated by their ability to restore sporulation competence to these cells. Three distinct plasmids were isolated. Each plasmid insert overlapped the same 8.0-kilobase region, and each plasmid restored ornithine decarboxylase activity to spe1A mutants. These plasmids also conferred ornithine decarboxylase activity to Escherichia coli EWH319 from which the ornithine decarboxylase gene is deleted. The plasmid-encoded activity expressed in E. coli resembled S. cerevisiae ornithine decarboxylase in its kinetic characteristics, indicating that the yeast ornithine decarboxylase gene was cloned. Southern blot analysis suggested that ornithine decarboxylase is a single-copy gene in S. cerevisiae. A single 2.1-kilobase transcript was demonstrated by Northern blot analysis.
机译：发现在不存在多胺的情况下，消除了鸟氨酸脱羧酶活性的spe1A突变的纯合酿酒酵母的二倍体细胞形成孢子。互补于spe1A突变的质粒通过其恢复这些细胞的孢子形成能力而被分离出来。分离出三个不同的质粒。每个质粒插入片段重叠相同的8.0千碱基区域，每个质粒对spe1A突变体恢复了鸟氨酸脱羧酶活性。这些质粒还赋予鸟氨酸脱羧酶活性给大肠杆菌EWH319，从其中删除了鸟氨酸脱羧酶基因。在大肠杆菌中表达的质粒编码活性在动力学特征上类似于酿酒酵母鸟氨酸脱羧酶，表明克隆了酵母鸟氨酸脱羧酶基因。 Southern印迹分析表明，鸟氨酸脱羧酶是酿酒酵母中的单拷贝基因。通过RNA印迹分析证实了单个2.1-碱基碱基的转录物。

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