Trezise A E, Cheung B, Holmes R S, Beacham I R
Division of Science and Technology, Griffith University, Nathan, Brisbane, Queensland, Australia.
Gene. 1991 Jul 22;103(2):211-8. doi: 10.1016/0378-1119(91)90275-g.
Five alcohol dehydrogenases (ADH; alcohol: NAD+ oxidoreductase; EC 1.1.1.1) have been identified in the baboon. All are homodimers of five distinct ADH subunits, with the two class-I ADH subunits being differentially expressed in the liver (the beta-subunit) and kidney. We have hybridized restriction-enzyme-digested baboon DNA to a 30-bp probe or a 337-bp DNA fragment, to reveal the presence of three genes encoding class-I ADH subunits in the baboon genome. This result was confirmed by the amplification of three different baboon ADH (bADH) nucleotide (nt) sequences, corresponding to exon 5 in the human gene encoding ADH beta (hADHB) from baboon DNA. Two of these sequences are identical to previously isolated liver and kidney cDNA nt sequences. These results are consistent with a phylogenetic analysis of the nt sequences of class-I hADH and bADH genes. Then, using primers based on the nt sequence of hADHB, we amplified a 336-bp DNA fragment, from genomic DNA, encoding the 5' region of the bADHB gene. In a 49-bp region of overlap, the nt sequence of this DNA fragment was identical to the sequence of a cDNA fragment amplified from baboon liver mRNA, whereas there were seven differences between this DNA fragment and the sequence of a cDNA amplified from baboon kidney mRNA. We used primer extension analysis to identify three adjacent transcriptional start points (tsp) for bADHB mRNA. Initiation of transcription at the most 5' bp leaves a 72-bp untranslated region. Examination of the sequence upstream from the tsp reveals a number of conserved putative regulatory sequence elements.
在狒狒中已鉴定出五种乙醇脱氢酶（ADH；乙醇：NAD+氧化还原酶；EC 1.1.1.1）。它们均为五个不同ADH亚基的同型二聚体，其中两个I类ADH亚基在肝脏（β亚基）和肾脏中差异表达。我们将经限制性酶切的狒狒DNA与一个30bp的探针或一个337bp的DNA片段进行杂交，以揭示狒狒基因组中存在三个编码I类ADH亚基的基因。通过从狒狒DNA中扩增出与人类ADHβ（hADHB）基因外显子5相对应的三个不同的狒狒ADH（bADH）核苷酸（nt）序列，证实了这一结果。其中两个序列与先前分离的肝脏和肾脏cDNA nt序列相同。这些结果与I类hADH和bADH基因的nt序列的系统发育分析一致。然后，我们根据hADHB的nt序列设计引物，从基因组DNA中扩增出一个336bp的DNA片段，该片段编码bADHB基因的5'区域。在一个49bp的重叠区域中，该DNA片段的nt序列与从狒狒肝脏mRNA扩增出的cDNA片段的序列相同，而该DNA片段与从狒狒肾脏mRNA扩增出的cDNA序列之间存在七个差异。我们使用引物延伸分析来确定bADHB mRNA的三个相邻转录起始点（tsp）。在最上游的bp处起始转录会留下一个72bp的非翻译区。对tsp上游序列的检查揭示了许多保守的假定调控序列元件。
