Feigenbaum A S, Robinson B H
Department of Genetics, University of Toronto, Ontario, Canada.
Genomics. 1993 Aug;17(2):376-81. doi: 10.1006/geno.1993.1335.
The structural organization of the gene for the E3 subunit of the human alpha-ketoacid dehydrogenase complexes, dihydrolipoamide dehydrogenase (DLD), and its upstream elements have been determined by restriction endonuclease mapping and DNA sequence analysis of overlapping genomic clones. The gene is approximately 20 kb long. It contains 14 exons ranging in size from 69 to 780 bp and 13 introns ranging in size from 93 bp to 7.0 kb. All splice donor and acceptor sites conform to the GT/AG rule. The 5' ends of mRNA transcripts upstream from the translation initiation codon were determined by primer extension assay. A "CAAT box"-like sequence is present at 39 bp upstream of the presumptive cap site and the 5' flanking region has been sequenced up to 2.0 kb upstream. There are several sequences compatible with presumptive promoter elements, including an Sp1 binding site, a nuclear respiratory factor 1 site, two cyclic AMP response element binding sites, and a possible negative response element present in the insulin promoter. A 313-bp segment from -2076 to -1763 is 89% homologous to a recently described pTR5 repetitive element found in the human genome.
通过对重叠基因组克隆进行限制性内切酶图谱分析和DNA序列分析，已确定了人类α-酮酸脱氢酶复合体E3亚基二氢硫辛酰胺脱氢酶（DLD）基因的结构组织及其上游元件。该基因长度约为20 kb。它包含14个外显子，大小从69 bp到780 bp不等，以及13个内含子，大小从93 bp到7.0 kb不等。所有剪接供体和受体位点均符合GT/AG规则。通过引物延伸试验确定了翻译起始密码子上游mRNA转录本的5'末端。在假定的帽位点上游39 bp处存在一个类似“CAAT盒”的序列，并且5'侧翼区域已测序至上游2.0 kb。有几个与假定启动子元件兼容的序列，包括一个Sp1结合位点、一个核呼吸因子1位点、两个环磷酸腺苷反应元件结合位点，以及胰岛素启动子中存在的一个可能的负反应元件。从-2076到-1763的一段313 bp序列与人类基因组中最近描述的pTR5重复元件有89%的同源性。
