Department of Life Science, Chang Gung University, Tao-Yuan, Taiwan.
Gene. 2010 May 1;455(1-2):16-21. doi: 10.1016/j.gene.2010.02.001. Epub 2010 Feb 10.
To meet the increasing need of simultaneously co-expressing two different genes in the same cell of transgenic Caenorhabditis elegans, here, we report the establishment of dicistronic vectors that contain an intercistronic region (ICR) of the C. elegans operon, CEOP5428. In these vectors, a green fluorescence protein (GFP) and a red FP (RFP) genes were placed in the first and second cistrons, respectively, which were separated by the ICR. Driven by the fibrillarin (fib-1) or myo-2 promoter, the GFP- and RFP-fusion proteins were consistently co-expressed in the entire worm cells or in the pharynx muscle cells of the transgenic worms, respectively. Our work demonstrates that ICR-containing dicistronic vectors could be developed into versatile co-expression systems in C. elegans for functional analysis of genes of interest.
为了满足在转基因秀丽隐杆线虫同一细胞中同时表达两种不同基因的日益增长的需求,我们在这里报告了含有秀丽隐杆线虫操纵子 CEOP5428 间插序列区(ICR)的双顺反子载体的建立。在这些载体中,绿色荧光蛋白(GFP)和红色荧光蛋白(RFP)基因分别置于第一顺反子和第二顺反子中,它们由 ICR 隔开。在纤毛蛋白(fib-1)或肌球蛋白-2 启动子的驱动下,GFP 和 RFP 融合蛋白分别在转基因蠕虫的整个蠕虫细胞或咽肌肉细胞中一致地共同表达。我们的工作表明,含有 ICR 的双顺反子载体可以在秀丽隐杆线虫中开发成多功能的共表达系统,用于感兴趣基因的功能分析。