Myasnikov A N, Sasnauskas K V, Janulaitis A A, Smirnov M N
Biological Institute, Leningrad University, U.S.S.R.
Gene. 1991 Dec 20;109(1):143-7. doi: 10.1016/0378-1119(91)90600-g.
The ADE1 gene of the yeast Saccharomyces cerevisiae has been cloned by complementation of the ade1 mutation. The nucleotide sequence has been determined for the 918-bp coding region, 240-bp 5'-noncoding region and 292-bp 3'-noncoding region. The sequenced region includes a single large open reading frame coding for a protein of 306 amino acid (aa) residues. The promoter of the ADE1 gene contains a copy of the 5'-TGACTC hexanucleotide, a feature characteristic of promoters under general aa control. Subsequent search of other published purine biosynthesis gene sequences revealed that all of them also contain general aa control signals in their promoter regions. An expression plasmid containing the ADE1 coding region under control of the PHO5 promoter produced N-succinyl-5-aminoimidazole-4-carboxamide ribotide (SAICAR) synthetase in yeast cells at a level of 40% of total cellular protein. One-step purification resulted in an almost homogeneous preparation of SAICAR synthetase.
通过对酿酒酵母ade1突变的互补作用，克隆了酿酒酵母的ADE1基因。已测定了918个碱基对的编码区、240个碱基对的5'非编码区和292个碱基对的3'非编码区的核苷酸序列。测序区域包括一个单一的大开放阅读框，编码一个由306个氨基酸残基组成的蛋白质。ADE1基因的启动子含有5'-TGACTC六核苷酸的一个拷贝，这是一般氨基酸控制下启动子的一个特征。随后对其他已发表的嘌呤生物合成基因序列的搜索表明，它们所有的启动子区域也都含有一般氨基酸控制信号。一个含有在PHO5启动子控制下的ADE1编码区的表达质粒，在酵母细胞中产生N-琥珀酰-5-氨基咪唑-4-甲酰胺核苷酸（SAICAR）合成酶，其水平占细胞总蛋白的40%。一步纯化得到了几乎纯的SAICAR合成酶制剂。
