Yeast thioredoxin genes.
Yeast thioredoxin genes.
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酵母硫氧还蛋白基因。
DOI：
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发表时间：
1991
影响因子：
4.8
通讯作者：
Z. Gan
中科院分区：
文献类型：
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作者：
Z. Gan
根据酵母和其他生物硫氧还蛋白的保守蛋白序列，合成了两条引物，用于酵母基因组DNA的聚合酶链反应。从聚合酶链反应产物中获得酵母硫氧还蛋白活性位点周围的34个碱基对（bp）序列。该特异性序列被用作探针，用于用各种限制性内切酶消化的总酵母基因组DNA的Southern印迹分析。在高严谨度条件下，多个DNA物种与探针杂交，表明多个基因编码酵母基因组文库。将来自两个不同克隆的两个Sau 3A 1片段，分别为825和2045 bp，克隆到pUC 13中。对这些片段的序列分析给出了两个不同的不含内含子的开放阅读框架。Sau 3A 1基因全长825 bp，编码一个含103个氨基酸残基的蛋白质，命名为硫氧还蛋白I。2045-bp的Sau 3A 1片段含有编码硫氧还蛋白II的序列，其具有102个氨基酸残基。这是第一次报道真核生物硫氧还蛋白基因的克隆和测序。两种酵母硫氧还蛋白都含有二巯基活性位点序列Cys-Gly-Pro-Cys。硫氧还蛋白I和II显示78%的氨基酸序列同一性。它们与哺乳动物硫氧还蛋白的氨基酸序列相似性比与大肠杆菌和植物叶绿体硫氧还蛋白的氨基酸序列相似性更高。
Based on the conserved protein sequence of thioredoxins from yeast and other organisms, two primers were synthesized for polymerase chain reaction of yeast genomic DNA. A 34-base pair (bp) sequence around the active site of yeast thioredoxin was obtained from the polymerase chain reaction product. This specific sequence was used as a probe in Southern blot analysis of total yeast genomic DNA digested with various restriction enzymes. Under conditions of high stringency, more than one DNA species hybridized with the probe, suggesting that more than one gene encodes yeast genomic library. Two Sau3A1 fragments, 825 and 2045 bp, respectively, from two different clones were cloned into pUC13. Sequence analysis of these fragments gave two different open reading frames without introns. The 825-bp Sau3A1 fragment encodes a 103-amino acid residue protein named thioredoxin I. The 2045-bp Sau3A1 fragment contains a sequence encoding thioredoxin II which has 102 amino acid residues. This is the first report of the cloning and sequencing of eukaryotic thioredoxin genes from any source. Both yeast thioredoxins contain a dithiol active site sequence, Cys-Gly-Pro-Cys. Thioredoxins I and II show 78% amino acid sequence identity. They display more amino acid sequence similarity with mammalian thioredoxin than with Escherichia coli and plant chloroplast thioredoxins.