Hygromycin resistance as a selectable marker in Dictyostelium discoideum.
Hygromycin resistance as a selectable marker in Dictyostelium discoideum.
复制标题
潮霉素抗性作为盘基网柄菌的选择标记。
DOI：
10.1128/mcb.9.5.1965-1968.1989
复制
发表时间：
1989
影响因子：
5.3
通讯作者：
Spudich,JA
中科院分区：
文献类型：
--
作者：
Egelhoff,TT;Brown,SS;Manstein,DJ;Spudich,JA
我们构建了一个表达盒，该表达盒将细菌潮霉素抗性基因（hph）融合到盘状网柄藻肌动蛋白15启动子上，并将肌动蛋白15位点的3′侧翼DNA片段置于hph基因下游作为转录终止子。将含有该盒和一个网柄藻复制起点的质粒pDE 109转化D.在潮霉素筛选条件下，这种选择性标记的可用性规避了先前将G418抗性作为该生物体的唯一选择性标记的限制;现在可以使用二次转化将DNA引入先前转化的细胞系中。
We have constructed an expression cartridge which has the bacterial hygromycin resistance gene (hph) fused to theDictyostelium discoideumactin 15 promoter, with a segment of 3′-flanking DNA from the actin 15 locus placed downstream of thehphgene to serve as a transcription terminator. The plasmid pDE109, which contained this cartridge and aDictyosteliumorigin of replication, transformedD. discoideumwith high efficiency under hygromycin selection. The availability of this selectable marker circumvents the previous limitation of having G418 resistance as the only selectable marker for this organism; secondary transformation can now be used to introduce DNA into previously transformed cell lines.