用绿色荧光蛋白基因作为筛选标记的新型克隆载体的构建

Construction of a Novel Cloning Vector and Screening the Recombinants by Green and White Colonies

用绿色荧光蛋白基因作为筛选标记的新型克隆载体的构建李寿东齐义鹏胡建红林宏（武汉大学病毒研究所武汉４３００７２）目前通常使用的质粒克隆载体，如ｐＵＣ系列、ｐＧＥＭ系列和ｐＢｌｕｅｓｃｒｉｐｔ等，都是以ｌａｃＺ’作为筛选标记的插入失活型载体。ｌａｃＺ’...

A novel cloning vector named pQLgreen was derived from pUC19 by insertion the ORF of gfp S65T gene into SmaI site of pUC19,under the control of LacZ promoter operator.Host bacteria transformed by pQLgreen may grow to blue colonies on Ap/IPTG LB plates.Green tag lost when the multiple restriction sites upstream the gfp S65T ORF was inserted by foreign DNA fragments.The colonies of host bacteria with recombinant plasmid showed colorless.So we incorporate a new colour selection for insertion,different from the blue and colourless selection system.It is economical because it does not require the use of costly X gal.