Altmann M, Handschin C, Trachsel H
Mol Cell Biol. 1987 Mar;7(3):998-1003. doi: 10.1128/mcb.7.3.998-1003.1987.
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.
我们已经分离出了编码酿酒酵母蛋白质合成起始因子eIF-4E(mRNA帽结合蛋白)的基因组和cDNA克隆。通过在大肠杆菌中表达cDNA确定了它们的身份。该cDNA编码的蛋白质在分子量、与m7GDP-琼脂糖亲和柱的结合及洗脱,以及蛋白水解肽谱方面与纯化的eIF-4E没有区别。通过将cDNA与酵母基因组文库的克隆进行杂交,分离出了eIF-4E基因。该基因没有内含子,单倍体基因组中每个基因组有一个拷贝,编码一个由213个氨基酸残基组成的蛋白质。基因破坏实验表明该基因对生长至关重要。