Kwak J W, Kim J, Yoo O J, Han M H
Genetic Engineering Center, KAIST, Seoul, Korea.
Gene. 1988 Apr 15;64(1):165-72. doi: 10.1016/0378-1119(88)90490-8.
A DNA fragment was isolated from yeast genomic sequences by its ability to direct the transcription of promoterless CmR (cat) gene in Escherichia coli and in yeast. Nucleotide sequencing and primer extension analysis showed that yeast DNA contains sets of consensus sequences pertaining to prokaryotic and yeast-type promoter elements. It was designated as yeast- and E. coli-type promoter (YEP1). Typical E. coli-type promoter elements are found at appropriate positions: TATTTT from -12 to -7 and TTGTCC from -35 to -30 with their spacing of 17 bp from the single mRNA start point determined by the primer extension. Analysis of cat transcripts from yeast cells showed that the YEP1 caused multiple transcription initiations at more than 20 different points that are spaced over a 100-bp region. The DNA is composed of A + T-rich sequences and putative TATA-like sequences are found at several places upstream from the transcription start points. Deletion analysis showed that a 276-bp sequence between -872 and -596 from the initiating ATG codon was required for the maximal promoter activity in yeast but not in E. coli.
通过其在大肠杆菌和酵母中指导无启动子CmR（氯霉素乙酰转移酶）基因转录的能力，从酵母基因组序列中分离出一个DNA片段。核苷酸测序和引物延伸分析表明，酵母DNA含有与原核生物和酵母型启动子元件相关的共有序列集。它被命名为酵母和大肠杆菌型启动子（YEP1）。在适当位置发现了典型的大肠杆菌型启动子元件：从-12到-7的TATTTT和从-35到-30的TTGTCC，通过引物延伸确定它们与单个mRNA起始点的间距为17 bp。对酵母细胞中氯霉素乙酰转移酶转录本的分析表明，YEP1在超过20个不同位点引发多个转录起始，这些位点分布在100 bp的区域内。该DNA由富含A + T的序列组成，并且在转录起始点上游的几个位置发现了推定的类TATA序列。缺失分析表明，从起始ATG密码子起-872至-596之间的276 bp序列对于酵母中的最大启动子活性是必需的，但在大肠杆菌中则不是。
