Hata A, Tsuzuki T, Shimada K, Takiguchi M, Mori M, Matsuda I
J Biochem. 1986 Sep;100(3):717-25. doi: 10.1093/oxfordjournals.jbchem.a121764.
We isolated over 20 phage clones carrying the ornithine transcarbamylase (OTC) [EC 2.1.3.3] gene, from two independently constructed human genomic DNA libraries, using as probes either a rat OTC cDNA or several nuclear DNA fragments derived from some of these isolated clones. These clones, classified into 10 different groups, overlapped and spanned a region of more than 85 kilobase pairs of the human genomic DNA. Restriction mapping and Southern blot analyses demonstrated that one of the clones covers the 5'-end region of the OTC gene. We sequenced the 5'-end region of the OTC gene and found that it covered 665 base pairs of the 5'-flanking region, the complete first exon and a part of the first intron (150 base pairs). In the 5'-flanking region, there were two pairs of putative CAAT and TATA boxes and one enhancer core-like sequence, GTGGAAAG. The first exon contained a coding region for most of the OTC presequence, i.e. 26 out of 32 amino acid residues of the presequence, including the initiation methionine.
我们从两个独立构建的人类基因组DNA文库中分离出20多个携带鸟氨酸转氨甲酰酶（OTC）[EC 2.1.3.3]基因的噬菌体克隆，分别使用大鼠OTC cDNA或从其中一些分离克隆中获得的几个核DNA片段作为探针。这些克隆分为10个不同的组，相互重叠并跨越了人类基因组DNA中超过85千碱基对的区域。限制性图谱分析和Southern印迹分析表明，其中一个克隆覆盖了OTC基因的5'端区域。我们对OTC基因的5'端区域进行了测序，发现它涵盖了665个碱基对的5'侧翼区域、完整的第一个外显子和第一个内含子的一部分（150个碱基对）。在5'侧翼区域，有两对推定的CAAT盒和TATA盒以及一个增强子核心样序列GTGGAAAG。第一个外显子包含大部分OTC前序列的编码区域，即前序列32个氨基酸残基中的26个，包括起始甲硫氨酸。
