Molecular and Cellular Biology · 1987 · 130 citations · 32 references
Life SciencesBiochemistryMolecular BiologyGene ExpressionGene FunctionProtein TranslationProtein SynthesisRna-binding ProteinsTranslation In EukaryotesTranslation In ProkaryotesTranslation InitiationTranslation RegulationCap Binding ProteinsCdna CloningCdna Library ConstructionEukaryotic Initiation Factor 4EEukaryotic Translation Initiation FactorsGene DisruptionMrna Translation InitiationProteolytic Profiles
We have isolated genomic and cDNA clones encoding protein synthesis initiation factor eIF-4E (mRNA cap-binding protein) of the yeast Saccharomyces cerevisiae. Their identity was established by expression of a cDNA in Escherichia coli. This cDNA encodes a protein indistinguishable from purified eIF-4E in terms of molecular weight, binding to and elution from m7GDP-agarose affinity columns, and proteolytic peptide pattern. The eIF-4E gene was isolated by hybridization of cDNA to clones of a yeast genomic library. The gene lacks introns, is present in one copy per haploid genome, and encodes a protein of 213 amino acid residues. Gene disruption experiments showed that the gene is essential for growth.
32
Molecular Cloning. A Laboratory Manual
EJ Wood · Biochemical Education · 1983 · 30.6K citations
Transformation of intact yeast cells treated with alkali cations
Hisao Ito, Y Fukuda, Kousaku Murata et al. · Journal of Bacteriology · 1983 · 7.7K citations · Full text