遗传 ›› 2004, Vol. 26 ›› Issue (4): 525-528.
刘 和1; 陈英旭1; 张文波2; 金勇丰2LIU He1; CHEN Ying-xu1; ZHANG Wen-bo2; JIN Yong-feng2
摘要:
采用一种不需要限制核酸酶和连接酶的新方法——“PCR一步法”将芳香烃化合物降解的关键基因pheB和绿色荧光蛋白编码基因gfp融合,构建得到融合蛋白基因fpg。该方法在一个PCR反应体系中通过三个引物、两个模板扩增得到一个含有中间柔性肽段-Gly4Ser-的融合基因fpg。本文研究结果表明,PCR一步法是一种快速方便的构建融合基因的方法。Abstract: TP-PCR,a method developed for fusion gene construction without the use of endonuclease and ligase, was performed to construct a fused fpg gene. The TP-PCR reaction system contained three primers and two templates and resulting PCR product, fused fpg gene, consisted of three sections: pheB gene, which was responsible for catechol 2,3-dioxygenase, gfp gene for GFP protein and the intermediate ligation segment which was designed for the correct expression of the fusion gene. The result in this paper showed that the TP-PCR method is one of rapid and convenient methods for fused gene construction.