Yu W, Macreadie I G, Winge D R
University of Utah Medical Center, School of Medicine, Salt Lake City 84132.
J Inorg Biochem. 1991 Nov 15;44(3):155-61. doi: 10.1016/0162-0134(91)80052-j.
A cDNA expression library from Schizosaccharomyces pombe was transformed into Saccharomyces cerevisiae to screen for genes capable of conferring cadmium resistance to S. cerevisiae cells. The cDNA library was cloned into the S. cerevisiae expression vector pDB20 which is designed to express cDNAs via the constitutively-expressed promoter of the gene for alcohol dehydrogenase I (ADH1). Terminator and polyadenylation signals are also provided by the ADH1 gene. Cadmium resistant colonies were shown to arise by a recombination event leading to the exchange of the S. pombe DNA with the chromosomal ADH1 gene and a consequent dramatic increase in the ADH1 gene expression due to the high copy number of the plasmid. The overexpression of ADH1 effectively buffered the cells for cadmium ions by formation of Cd-ADH.
将粟酒裂殖酵母的一个cDNA表达文库转化到酿酒酵母中,以筛选能够赋予酿酒酵母细胞镉抗性的基因。该cDNA文库被克隆到酿酒酵母表达载体pDB20中,该载体设计用于通过组成型表达的乙醇脱氢酶I(ADH1)基因的启动子来表达cDNA。ADH1基因还提供终止子和聚腺苷酸化信号。结果表明,镉抗性菌落是通过重组事件产生的,该事件导致粟酒裂殖酵母DNA与染色体ADH1基因发生交换,并且由于质粒的高拷贝数,ADH1基因表达随之急剧增加。ADH1的过表达通过形成Cd-ADH有效地缓冲了细胞对镉离子的影响。