Cloning and analysis of a yeast genomic DNA sequence capable of directing gene transcription in Escherichia coli as well as in yeast.
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.
Kwak
J W
JW
Genetic Engineering Center, KAIST, Seoul, Korea.
Kim
J
J
Yoo
O J
OJ
Han
M H
MH
D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004271:DNA, Fungal / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D005798:Genes, Bacterial; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics*
D004271:DNA, Fungal; D004262:DNA Restriction Enzymes