Ceci J D, Lawther R, Duester G, Hatfield G W, Smith M, O'Malley M P, Felder M R
Gene. 1986;41(2-3):217-24. doi: 10.1016/0378-1119(86)90101-0.
A cDNA clone for the beta-chain of human alcohol dehydrogenase (ADH) was used to isolate several cross-hybridizing clones from a mouse liver cDNA library. Clones pADHm9 and a portion of pADHm12 were sequenced. pADHm9 coded for a sequence of 151 C-terminal amino acids and some untranslated sequences from the 3' end of its corresponding mRNA. This clone was identified as an Adh-1 cDNA clone. Consistent with the known expression of Adh-1, this gene was expressed constitutively in liver, whereas the Adh-3 gene product was found only in stomach, lung and reproductive tissues. Furthermore, the translated region of the cDNA shared 91% amino acid sequence homology with rat liver ADH. [32P]pADHm9 was used as a hybridization probe to study the mechanism of androgen induction of kidney ADH activity. Induction of A/J female mice by androgen resulted in a dramatic increase in the steady-state level of Adh-1 mRNA content which correlated with the level of enzyme induction. The size of the mRNA obtained from control or induced kidney and liver tissues was indistinguishable by Northern analysis. [32P]pADHm9 was also used to probe restriction fragments of genomic DNA obtained from several inbred mouse strains. The hybridization patterns, considered with the genetic evidence, suggested that pADHm9 recognized sequences which may be present as only a single copy in the genome. No restriction fragment length polymorphisms were observed among the several inbred mouse strains examined.
利用人乙醇脱氢酶（ADH）β链的cDNA克隆从小鼠肝脏cDNA文库中分离出几个交叉杂交克隆。对克隆pADHm9和pADHm12的一部分进行了测序。pADHm9编码其相应mRNA 3'端的151个C末端氨基酸序列和一些非翻译序列。该克隆被鉴定为Adh-1 cDNA克隆。与已知的Adh-1表达一致，该基因在肝脏中组成性表达，而Adh-3基因产物仅在胃、肺和生殖组织中发现。此外，该cDNA的翻译区域与大鼠肝脏ADH的氨基酸序列同源性为91%。[32P]pADHm9用作杂交探针研究雄激素诱导肾脏ADH活性的机制。雄激素诱导A/J雌性小鼠导致Adh-1 mRNA含量的稳态水平显著增加，这与酶诱导水平相关。通过Northern分析，从对照或诱导的肾脏和肝脏组织中获得的mRNA大小无法区分。[32P]pADHm9还用于探测从几个近交小鼠品系获得的基因组DNA的限制性片段。结合遗传证据考虑，杂交模式表明pADHm9识别的序列可能在基因组中仅以单拷贝形式存在。在所检测的几个近交小鼠品系中未观察到限制性片段长度多态性。
