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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.
Author(s) -
Nagata S.,
Nagashima K.,
TsunetsuguYokota Y.,
Fujimura K.,
Miyazaki M.,
Kaziro Y.
Publication year - 1984
Publication title -
the embo journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 7.484
H-Index - 392
eISSN - 1460-2075
pISSN - 0261-4189
DOI - 10.1002/j.1460-2075.1984.tb02053.x
Subject(s) - biology , saccharomyces cerevisiae , alpha (finance) , eukaryotic translation elongation factor 1 alpha 1 , nucleic acid sequence , genetics , elongation factor , yeast , gene , alpha chain , nucleotide , microbiology and biotechnology , rna , medicine , ribosome , nursing , patient satisfaction , construct validity
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.