摘要
利用质粒YEplac112为骨架,在其多克隆位点Hind III和EcoRI之间插入水母绿色荧光蛋白(GFP)开放阅读框上下游约1 kb左右的DNA序列,得到YEplac112-CT质粒,在其多克隆位点Pst I和Bam HI之间插入750bp左右的GFP DNA序列,构建表达质粒YEplac112-C-GFP-T,以酿酒酵母W303为宿主,通过报告基因GFP验证了所构建的表达载体具有便捷筛选及良好的表达效果。
In this paper,plasmid YEplac112 skeleton was used,green fluorescent protein ( GFP ) was inserted in the multiple cloning sites between HindIII and EcoRI in the open reading frame downstream about 1 KB of DNA sequence, plasmid YEplac112-CT was obtained. 750bp left by the GFP DNA sequence was inserted in the multiple cloning sites PstI and BamHI. Using Saccharomyces cerevisiae W303 as the host, it is proved by reporter gene GFP that the constructed expression vector has good effect,convenient screening and expression.
出处
《齐鲁工业大学学报》
2016年第3期24-27,共4页
Journal of Qilu University of Technology
基金
国家自然科学资金(31370110)
关键词
绿色荧光蛋白
GFP
质粒
表达
green fluorescent protein
GFP
plasmid
expression