Webster C I, Brammar W J
ICI/University Joint Laboratory, Department of Biochemistry, University of Leicester, UK.
Microbiology (Reading). 1995 Sep;141 ( Pt 9):2191-200. doi: 10.1099/13500872-141-9-2191.
The ability of the bacteriophage 434 operator/repressor system to function in a eukaryotic cell has been explored. An idealized 434 operator was placed at various positions in the PGK promoter of Saccharomyces cerevisiae: within the upstream activator sequence, close to the TATA box, and downstream of the transcription-initiation site. Expression of the 434 cI gene from a 2 microns-based plasmid resulted in significant repression of gene expression from constructs containing the altered promoters linked to a beta-galactosidase reporter gene. Attempts to use a variant of the 434 repressor that has the binding specificity of the P22 repressor (434P22) were unsuccessful, due to a severely inhibitory effect of this gene-product on the growth of the yeast cells.
噬菌体434操纵子/阻遏物系统在真核细胞中发挥功能的能力已得到研究。一个理想化的434操纵子被置于酿酒酵母PGK启动子的不同位置:在上游激活序列内、靠近TATA框以及转录起始位点下游。从基于2μm的质粒表达434 cI基因,导致含有与β-半乳糖苷酶报告基因相连的改变启动子的构建体的基因表达受到显著抑制。由于该基因产物对酵母细胞生长具有严重抑制作用,尝试使用具有P22阻遏物结合特异性的434阻遏物变体(434P22)未成功。