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酿酒酵母GAL7基因的一级结构。

Primary structure of the Saccharomyces cerevisiae GAL7 gene.

作者信息

Tajima M, Nogi Y, Fukasawa T

机构信息

Laboratory of Molecular Genetics, Keio University School of Medicine, Tokyo, Japan.

出版信息

Yeast. 1985 Sep;1(1):67-77. doi: 10.1002/yea.320010108.

Abstract

We present the nucleotide sequence of a 1599-base pair (bp) DNA fragment containing the entire GAL7 gene that encodes galactose-1-phosphate uridyltransferase of Saccharomyces cerevisiae. The deduced peptide was composed of 364 amino acid residues. The expected molecular weight was 42,005 daltons, which agreed with the observed value for the purified enzyme. The 3'-end of the GAL7 transcript mapped at a position 82 bp downstream from the UAA termination codon by the S1 nuclease protection experiment. We constructed a GAL7'-lac'Z fusion on various types of yeast plasmid vectors. The fused gene on any type of vector was induced by galactose and repressed by glucose as for the GAL7 gene on the chromosome. The response of GAL7'-lac'Z fusion to gal4 delta and gal80 delta regulatory mutations was also similar to the response of the chromosomal GAL7 gene. By using various deletions in the 5'-flanking region of the gene fusion, we delimited the sequence essential for galactose controlled expression with a 180 bp-fragment of DNA lying 92 bp upstream of the transcription initiation site.

摘要

我们展示了一个1599个碱基对(bp)的DNA片段的核苷酸序列,该片段包含编码酿酒酵母半乳糖-1-磷酸尿苷转移酶的完整GAL7基因。推导的肽由364个氨基酸残基组成。预期分子量为42,005道尔顿,这与纯化酶的观察值一致。通过S1核酸酶保护实验,GAL7转录本的3'端定位在UAA终止密码子下游82 bp处。我们在各种类型的酵母质粒载体上构建了GAL7'-lac'Z融合体。与染色体上的GAL7基因一样,任何类型载体上的融合基因都受半乳糖诱导并受葡萄糖抑制。GAL7'-lac'Z融合体对gal4δ和gal80δ调控突变的反应也与染色体GAL7基因的反应相似。通过在基因融合体的5'侧翼区域进行各种缺失操作,我们用位于转录起始位点上游92 bp处的180 bp DNA片段确定了对半乳糖控制表达至关重要的序列。

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