论文部分内容阅读
采用巢式PCR获得甘蓝SCR的成熟肽编码区,利用同源重组技术首次将其克隆到pGBKT7载体中,构建酵母双杂交系统的诱饵载体pGBKT7-SCR,结果表明:通过巢式PCR获得了正确的甘蓝SCR成熟肽编码区,并成功构建到pGBKT7诱饵载体中,且转化有诱饵载体的Y2HGold在SD/-Trp营养缺陷平板上生长良好,而在SD/-His-Trp和SD/-Trp/X-a-Gal/AbA营养缺陷平板上皆不能生长,说明对报告基因无自激活作用;且毒性实验也表明,SCR蛋白对酵母没有毒害作用.
The mature peptide coding region of SCR was obtained by nested PCR and cloned into pGBKT7 vector using homologous recombination technology. The bait vector pGBKT7-SCR of yeast two-hybrid system was constructed. The results showed that the correct Brassica napus mature peptide coding region and was successfully constructed into pGBKT7 bait vector, and Y2HGold transformed with bait vector grew well on SD / -Trp auxotrophic plate, while SD / -His-Trp and SD / -Trp / Xa - Gal / AbA auxotrophy can not grow on the plate, indicating no self-activation of the reporter gene; and toxicity experiments also showed that the SCR protein has no toxic effects on yeast.