Cloning, nucleotide sequence, and expression of one of two genes coding for yeast elongation factor 1 alpha.
One gene coding for yeast cytoplasmic elongation factor 1 alpha (EF-1 alpha) was isolated by colony hybridization using a cDNA probe prepared from purified EF-1 alpha mRNA. A recombinant plasmid, pLB1, with a 6-kilobase yeast DNA insert, was found by hybrid selection and translation experiments to carry the entire gene. The nucleotide sequence of the gene with its 5'- and 3'-flanking regions was determined. The 5' and 3' ends of EF-1 alpha mRNA were localized by the S1 nuclease mapping technique. The cloned gene, called TEF1, encodes a protein of 458 amino acids (Mr = 50,071) in a single, uninterrupted reading frame. The amino acid sequence shows a strong homology with several domains of Artemia salina EF-1 alpha cytoplasmic factor, as evidenced by diagonal dot matrix analysis. Protein sequence homology is comparatively much lower with the yeast mitochondrial elongation factor. S1 nuclease mapping of the mRNA, hybridization analysis of chromosomal DNA using intragenic or extragenic DNA probes, and gene disruption experiments demonstrated the existence of two genes coding for the cytoplasmic elongation factor EF-1 alpha/haploid genome. The presence of an intact chromosomal TEF1 gene is not essential for growth of haploid yeast cells.
Cottrelle
P
P
Thiele
D
D
Price
V L
VL
Memet
S
S
Micouin
J Y
JY
Marck
C
C
Buhler
J M
JM
Sentenac
A
A
Fromageot
P
P
D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004247:DNA / Q000032:analysis; D004262:DNA Restriction Enzymes / Q000378:metabolism; D004720:Endonucleases / Q000378:metabolism; D005786:Gene Expression Regulation; D009693:Nucleic Acid Hybridization; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D015719:Single-Strand Specific DNA and RNA Endonucleases
D003062:Codon; D020648:Peptide Elongation Factor 1; D010445:Peptide Elongation Factors; D004247:DNA; D004720:Endonucleases; D004262:DNA Restriction Enzymes; D015719:Single-Strand Specific DNA and RNA Endonucleases