Polypeptide chain elongation factor 1 alpha (EF‐1 alpha) from yeast: nucleotide sequence of one of the two genes for EF‐1 alpha from Saccharomyces cerevisiae.
Polypeptide chain elongation factor 1 alpha (EF‐1 alpha) from yeast: nucleotide sequence of one of the two genes for EF‐1 alpha from Saccharomyces cerevisiae.
复制标题
来自酵母的多肽链延伸因子 1 α (EF-1 α)：来自酿酒酵母的 EF-1 α 的两个基因之一的核苷酸序列。
作者：
S. Nagata;K. Nagashima;Y. Tsunetsugu;K. Fujimura;M. Miyazaki;Y. Kaziro
从酿酒酵母中部分纯化了酵母胞质多肽链延伸因子1 α （EF‐1 α）的信使RNA。以pBR327为载体，在大肠杆菌中合成并克隆了双链互补DNA （cDNA）。在非严格条件下，将携带yEF‐1 α cDNA的重组质粒与大肠杆菌tufB基因和酵母线粒体EF‐Tu基因（tufM）进行交叉杂交鉴定。然后用EF‐1 α cDNA筛选酵母基因文库，并分离出几个含有EF‐1 α染色体基因的克隆。对这些克隆的DNA片段进行限制性内切分析，并对酵母基因组DNA与标记的EF‐1 α cDNA进行Southern杂交，结果表明酿酒酵母中存在两个EF‐1 α基因。两个EF‐1 α基因之一的核苷酸序列（命名为EF1 α A）及其5′‐和3′‐侧链序列被建立。该序列包含1374个核苷酸，编码458个氨基酸的蛋白质，计算的分子量为50 300。所得氨基酸序列与酵母线粒体EF‐Tu和大肠杆菌EF‐Tu的同源性分别为31%和32%。
Messenger RNA for yeast cytosolic polypeptide chain elongation factor 1 alpha (EF‐1 alpha) was partially purified from Saccharomyces cerevisiae. Double‐stranded complementary DNA (cDNA) was synthesized and cloned in Escherichia coli with pBR327 as a vector. Recombinant plasmid carrying yEF‐1 alpha cDNA was identified by cross‐hybridization with the E. coli tufB gene and the yeast mitochondrial EF‐Tu gene (tufM) under non‐stringent conditions. A yeast gene library was then screened with the EF‐1 alpha cDNA and several clones containing the chromosomal gene for EF‐1 alpha were isolated. Restriction analysis of DNA fragments of these clones as well as the Southern hybridization of yeast genomic DNA with labelled EF‐1 alpha cDNA indicated that there are two EF‐1 alpha genes in S. cerevisiae. The nucleotide sequence of one of the two EF‐1 alpha genes (designated as EF1 alpha A) was established together with its 5′‐ and 3′‐flanking sequences. The sequence contained 1374 nucleotides coding for a protein of 458 amino acids with a calculated mol. wt. of 50 300. The derived amino acid sequence showed homologies of 31% and 32% with yeast mitochondrial EF‐Tu and E. coli EF‐Tu, respectively.