Cloning, nucleotide sequence, and expression of one of two genes coding for yeast elongation factor 1 alpha.

摘要: 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 mRNA. A recombinant plasmid, pLB1, with 6-kilobase DNA insert, found hybrid selection and translation experiments to carry the entire gene. The nucleotide sequence of its 5'- 3'-flanking regions determined. 5' 3' ends mRNA were localized S1 nuclease mapping technique. cloned gene, called TEF1, encodes protein 458 amino acids (Mr = 50,071) in single, uninterrupted reading frame. acid shows strong homology several domains Artemia salina factor, as evidenced diagonal dot matrix analysis. Protein is comparatively much lower mitochondrial factor. mRNA, analysis chromosomal intragenic or extragenic probes, disruption demonstrated existence two genes alpha/haploid genome. presence an intact TEF1 not essential growth haploid cells.