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线虫秀丽隐杆线虫中的甘油醛-3-磷酸脱氢酶基因家族:其中一个基因的分离与鉴定

The glyceraldehyde-3-phosphate dehydrogenase gene family in the nematode, Caenorhabditis elegans: isolation and characterization of one of the genes.

作者信息

Yarbrough P O, Hayden M A, Dunn L A, Vermersch P S, Klass M R, Hecht R M

出版信息

Biochim Biophys Acta. 1987 Jan 28;908(1):21-33. doi: 10.1016/0167-4781(87)90018-2.

DOI:10.1016/0167-4781(87)90018-2
PMID:3026481
Abstract

The isolation and genomic sequence of one of possibly four glyceraldehyde-3-phosphate dehydrogenase genes in the nematode, Caenorhabditis elegans is presented. The complete nucleotide sequence of the coding as well as the noncoding flanking regions of this gene has been determined. The deduced amino-acid sequence agrees with the sequence of typical glyceraldehyde-3-phosphate dehydrogenase enzymes and its molecular weight of 36,235 agrees with its size determined previously (Yarbrough, P. and Hecht, R. (1984) J. Biol. Chem. 259, 14711-14720). That this isolated gene encodes a nematode glyceraldehyde-3-phosphate dehydrogenase is additionally confirmed by demonstrating its immunoreactivity to an anti-nematode glyceraldehyde-3-phosphate dehydrogenase antibody after its expression as a fusion protein with dihydrofolate reductase. Codon utilization follows a pattern typical of other expressed nematode genes. The gene is split by two introns that are highly conserved in comparison to other introns observed in C. elegans. The placement of one of these introns is conserved with respect to the chicken glyceraldehyde-3-phosphate dehydrogenase gene. Within the 5' flanking sequence homology to actin and the homology 2 block of the major myosin gene (unc-54) is noted. It is of interest that the 3' flanking region contains a CAAAT box, followed by a TATAAT box, before an open reading frame of a closely linked gene that also contains a small AT-rich intron with the nematode consensus splice junction.

摘要

本文介绍了线虫秀丽隐杆线虫中可能存在的四个3-磷酸甘油醛脱氢酶基因之一的分离及基因组序列。已确定该基因编码区以及非编码侧翼区的完整核苷酸序列。推导的氨基酸序列与典型的3-磷酸甘油醛脱氢酶的序列一致,其分子量为36235,与先前确定的大小相符(亚尔布罗,P.和赫希特,R.(1984年)《生物化学杂志》259,14711 - 14720)。通过将其作为与二氢叶酸还原酶的融合蛋白表达后,证明其与抗线虫3-磷酸甘油醛脱氢酶抗体具有免疫反应性,进一步证实了这个分离的基因编码线虫的3-磷酸甘油醛脱氢酶。密码子使用遵循其他已表达的线虫基因的典型模式。该基因被两个内含子隔开,与在秀丽隐杆线虫中观察到的其他内含子相比,这两个内含子高度保守。其中一个内含子的位置与鸡的3-磷酸甘油醛脱氢酶基因是保守的。在5'侧翼序列中,注意到与肌动蛋白的同源性以及主要肌球蛋白基因(unc-54)的同源性2区。有趣的是,3'侧翼区包含一个CAAAT框,接着是一个TATAAT框,然后是一个紧密相连基因的开放阅读框,该基因也包含一个带有线虫共有剪接位点的富含AT的小内含子。

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