生物
乌拉3
多克隆站点
克隆(编程)
遗传学
质粒
克隆载体
终端(太阳能)
基因
酿酒酵母
表达式向量
发起人
分子克隆
异源表达
互补DNA
分子生物学
基因表达
计算生物学
重组DNA
计算机科学
电离层
物理
程序设计语言
天文
作者
Dominik Mumberg,Rolf Müller,Martin Funk
出处
期刊:Gene
[Elsevier]
日期:1995-04-01
卷期号:156 (1): 119-122
被引量:1828
标识
DOI:10.1016/0378-1119(95)00037-7
摘要
An expression system for Saccharomyces cerevisiae (Sc) has been developed which, depending on the chosen vector, allows the constitutive expression of proteins at different levels over a range of three orders of magnitude and in different genetic backgrounds. The expression system is comprised of cassettes composed of a weak CYC1 promoter, the ADH promoter or the stronger TEF and GPD promoters, flanked by a cloning array and the CYC1 terminator. The multiple cloning array based on pBIISK (Stratagene) provides six to nine unique restriction sites, which facilitates the cloning of genes and allows for the directed cloning of cDNAs by the widely used ZAP system (Stratagene). Expression cassettes were placed into both the centromeric and 2μ plasmids of the pRS series [Sikorski and Hieter, Genetics 122 (1989) 19-27; Christianson et al., Gene 110 (1992) 119-122] containing HIS3, TRP1, LEU2 or URA3 markers. The 32 expression vectors created by this strategy provide a powerful tool for the convenient cloning and the controlled expression of genes or cDNAs in nearly every genetic background of the currently used Sc strains.
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