Holland J P, Holland M J
J Biol Chem. 1980 Mar 25;255(6):2596-605.
A hybrid plasmid (pgap63) was isolated which contains a second yeast glyceraldehyde-3-phosphate dehydrogenase structural gene. The complete nucleotide sequence of this gene was determined and compared with the primary structure of a yeast glyceraldehyde-3-phosphate dehydrogenase gene (pgap49) which was reported previously (Holland, J.P., and Holland, M.J. (1979) J. Biol. Chem. 254, 9839-9845). Based on the restriction endonuclease cleavage maps of the isolated segments of yeast DNA which contain these genes, the two genes are nontandemly duplicated. Greater than 94% of the nucleotides within the coding regions of these genes are homologous and the polypeptides encoded by the two structural genes differ by only 15 amino acid residues. Both genes have the same, highly biased, codon usage pattern and neither contains intervening sequences. Approximately 100 nucleotides adjacent to the ATG initiation codons and 130 nucleotides beyond the TAA termination codons are greater than 70% homologous. Structures within the flanking sequences of the genes which are potentially relevant to transcriptional and translational control are described. Several sequences (8 to 15 nucleotides in length) are repeated in both the 5' and 3' flanking sequences of the genes in a noninverted fashion. Finally, a rapid procedure for the isolation of spontaneous deletions within hybrid plasmid DNAs is described, as is the isolation of a structural gene deletion in pgap49.
分离出一种杂种质粒(pgap63),它含有第二个酵母甘油醛-3-磷酸脱氢酶结构基因。测定了该基因的完整核苷酸序列,并与先前报道的酵母甘油醛-3-磷酸脱氢酶基因(pgap49)的一级结构进行了比较(霍兰德,J.P.,和霍兰德,M.J.(1979年)《生物化学杂志》254,9839 - 9845)。根据含有这些基因的酵母DNA分离片段的限制性内切酶切割图谱,这两个基因是非串联重复的。这些基因编码区内超过94%的核苷酸是同源的,两个结构基因编码的多肽仅相差15个氨基酸残基。两个基因具有相同的、高度偏向性的密码子使用模式,且都不包含间隔序列。与ATG起始密码子相邻的大约100个核苷酸以及TAA终止密码子之后的130个核苷酸的同源性大于70%。描述了基因侧翼序列中可能与转录和翻译控制相关的结构。几个序列(长度为8至15个核苷酸)以非反向方式在基因的5'和3'侧翼序列中重复出现。最后,描述了一种快速分离杂种质粒DNA中自发缺失的方法,以及pgap49中一个结构基因缺失的分离方法。