Martins A S, Greene L J, Yoho L L, Milsted A
Department of Brain and Vascular Research, Cleveland Clinic Foundation, OH 44195, USA.
Gene. 1995 Aug 19;161(2):253-7. doi: 10.1016/0378-1119(95)00268-b.
A 2288-bp cDNA sequence encoding dihydrolipoamide dehydrogenase (DLDH; dihydrolipoamide: NAD+ oxido-reductase; EC 1.8.1.4) was obtained by isolating a 1762-bp cDNA clone from a canine skeletal muscle library in the vector, lambda UNIZAP, combined with PCR amplification of the 5' end of the mRNA. The DLDH cDNA sequence contains a 49-bp G+C-rich 5'-untranslated region (UTR), followed by 1527 bp of coding region, and 695 bp of 3'-UTR preceding a 17-bp poly(A) tail. The single open reading frame encodes a precursor DLDH of 509 amino acids (aa) that begins with a 35-aa leader sequence. The 3'-UTR includes six possible polyadenylation signals (three AATAAA, one TATAAA and two AATGAA) and one potential stem-loop region extending from bp 1969-1991. Alignment studies of the canine and human DLDH demonstrate homology within the coding region of 98% at the aa level and 94% at the nt level. Northern blot analysis using the cDNA clone as probe showed wide tissue distribution of the mRNA, with differences in the level of expression among tissues and possible utilization of different polyadenylation sites.
通过从载体λUNIZAP中的犬骨骼肌文库中分离出一个1762 bp的cDNA克隆,并结合mRNA 5'端的PCR扩增,获得了一个编码二氢硫辛酰胺脱氢酶(DLDH;二氢硫辛酰胺:NAD+氧化还原酶;EC 1.8.1.4)的2288 bp cDNA序列。DLDH cDNA序列包含一个49 bp富含G+C的5'非翻译区(UTR),随后是1527 bp的编码区,以及在17 bp多聚腺苷酸尾巴之前的695 bp 3'-UTR。单一开放阅读框编码一个509个氨基酸(aa)的前体DLDH,其起始于一个35个氨基酸的前导序列。3'-UTR包括六个可能的聚腺苷酸化信号(三个AATAAA、一个TATAAA和两个AATGAA)以及一个从第1969至1991碱基对延伸的潜在茎环区域。犬和人DLDH的比对研究表明,编码区在氨基酸水平上的同源性为98%,在核苷酸水平上为94%。使用该cDNA克隆作为探针的Northern印迹分析显示,该mRNA在组织中广泛分布,各组织间表达水平存在差异,且可能利用了不同的聚腺苷酸化位点。