Hsu L C, Tani K, Fujiyoshi T, Kurachi K, Yoshida A
Proc Natl Acad Sci U S A. 1985 Jun;82(11):3771-5. doi: 10.1073/pnas.82.11.3771.
Partial cDNA clones encoding human cytosolic aldehyde dehydrogenase (ALDH1) and mitochondrial aldehyde dehydrogenase (ALDH2) were isolated from a human liver cDNA library constructed in phage lambda gt11. The expression library was screened by using rabbit antibodies against ALDH1 and ALDH2. Positive clones thus obtained were subsequently screened with mixed synthetic oligonucleotides compatible with peptide sequences of ALDH1 and ALDH2. One of the positive clones for ALDH1 contained an insertion of 1.6 kilobase pairs (kbp). The insert encoded 340 amino acid residues and had a 3' noncoding region of 538 bp and a poly(A) segment. The amino acid sequence deduced from the cDNA sequence coincided with the reported amino acid sequence of human ALDH1 [Hempel, J., von Bahr-Lindström, H. & Jörnvall, H. (1984) Eur. J. Biochem. 141, 21-35], except that valine at position 161 in the previous amino acid sequence study was found to be isoleucine in the deduced sequence. Since the amino acid sequence of ALDH2 was unknown, 33 tryptic peptides of human ALDH2 were isolated and sequenced. Based on the amino acid sequence data thus obtained, a mixed oligonucleotide probe was prepared. Two positive clones, lambda ALDH2-21 and lambda ALDH2-36, contained the same insert of 1.2 kbp. Another clone, lambda ALDH2-22, contained an insert of 1.3 kbp. These two inserts contained an overlap region of 0.9 kbp. The combined cDNA contained a sequence that encodes 399 amino acid residues, a chain-termination codon, a 3' untranslated region of 403 bp, and a poly(A) segment. The deduced amino acid sequence was compatible with the amino acid sequences of the tryptic peptides. The degree of homology between human ALDH1 and ALDH2 is 66% for the coding regions of their cDNAs and 69% at the protein level. No significant homology was found in their 3' untranslated regions.
从构建于噬菌体λgt11中的人肝脏cDNA文库中分离出编码人胞质醛脱氢酶（ALDH1）和线粒体醛脱氢酶（ALDH2）的部分cDNA克隆。使用针对ALDH1和ALDH2的兔抗体筛选该表达文库。随后用与ALDH1和ALDH2肽序列兼容的混合合成寡核苷酸对由此获得的阳性克隆进行筛选。ALDH1的一个阳性克隆含有一个1.6千碱基对（kbp）的插入片段。该插入片段编码340个氨基酸残基，具有一个538 bp的3'非编码区和一个聚腺苷酸（poly(A)）片段。从cDNA序列推导的氨基酸序列与报道的人ALDH1氨基酸序列一致[亨佩尔，J.，冯·巴尔-林德斯特伦，H.和约恩瓦尔，H.（1984年）《欧洲生物化学杂志》141，21 - 35]，只是在先前氨基酸序列研究中第161位的缬氨酸在推导序列中被发现为异亮氨酸。由于ALDH2的氨基酸序列未知，分离并测序了人ALDH2的33个胰蛋白酶肽段。基于由此获得的氨基酸序列数据，制备了混合寡核苷酸探针。两个阳性克隆，λALDH2 - 21和λALDH2 - 36，含有相同的1.2 kbp插入片段。另一个克隆，λALDH2 - 22，含有一个1.3 kbp的插入片段。这两个插入片段包含一个0.9 kbp的重叠区域。组合的cDNA包含一个编码399个氨基酸残基的序列、一个链终止密码子、一个4
