Ogawa H, Miller D A, Dunn T, Su Y, Burcham J M, Peraino C, Fujioka M, Babcock K, Pitot H C
McArdle Laboratory for Cancer Research, Medical School, University of Wisconsin, Madison 53706.
Proc Natl Acad Sci U S A. 1988 Aug;85(16):5809-13. doi: 10.1073/pnas.85.16.5809.
Rat serine dehydratase cDNA clones were isolated from a lambda gt11 cDNA library on the basis of their reactivity with monospecific immunoglobulin to the purified enzyme. Using the cDNA insert from a clone that encoded the serine dehydratase subunit as a probe, additional clones were isolated from the same library by plaque hybridization. Nucleotide sequence analysis of the largest clone obtained showed that it has 1444 base pairs with an open reading frame consisting of 1089 base pairs. The deduced amino acid sequence contained sequences of several portions of the serine dehydratase protein, as determined by Edman degradation. Rat liver serine dehydratase mRNA virtually disappeared from livers of rats fed a protein-free diet for 5 days. Several genomic clones were isolated from two libraries. Determinations of the transcription start site and the structure of the 3' flanking region of the gene indicated that the coded mRNA is 1504 nucleotides long. The 5' promoter region contained a variety of sequences similar to several consensus sequences believed to be important for the regulation of specific gene expression.
大鼠丝氨酸脱水酶cDNA克隆是基于其与针对纯化酶的单特异性免疫球蛋白的反应性，从λgt11 cDNA文库中分离得到的。使用来自编码丝氨酸脱水酶亚基的克隆的cDNA插入片段作为探针，通过噬菌斑杂交从同一文库中分离出其他克隆。对获得的最大克隆进行核苷酸序列分析表明，它有1444个碱基对，开放阅读框由1089个碱基对组成。推导的氨基酸序列包含丝氨酸脱水酶蛋白几个部分的序列，这是通过埃德曼降解法确定的。给大鼠喂食无蛋白饮食5天后，大鼠肝脏中的丝氨酸脱水酶mRNA几乎消失。从两个文库中分离出了几个基因组克隆。对该基因转录起始位点和3'侧翼区域结构的测定表明，编码的mRNA长1504个核苷酸。5'启动子区域包含多种与一些被认为对特定基因表达调控很重要的共有序列相似的序列。
