橡胶草3-羟基-3-甲基戊二酰辅酶A基因表达的初步分析

发布时间：2018-04-25 04:41

  本文选题：橡胶草 + HMGR基因　； 参考：《江苏农业科学》2017年16期

【摘要】：分析橡胶草HMGR基因的表达情况,构建原核表达载体,转化BL21菌株并诱导表达。构建细胞融合表达载体,转化GV3101菌株并侵染洋葱表皮细胞,在共聚焦显微镜下观察基因表达情况。构建植物过表达载体并侵染烟草,利用紫外分光光度法检测三萜含量的变化。通过IPTG诱导和SDS-PAGE检测,获得分子量为62.659 1 ku的蛋白,与预期蛋白相符合,且37℃诱导6 h时表达最明显;通过激光共聚焦显微镜观察,该基因主要在细胞膜上表达;紫外可见分光光度法表明转基因烟草中三萜的含量高于空白烟草。橡胶草HMGR基因分别在大肠杆菌、洋葱鳞片内表皮和烟草中成功表达。
[Abstract]:The expression of HMGR gene was analyzed, the prokaryotic expression vector was constructed, BL21 strain was transformed and induced expression. Cell fusion expression vector was constructed, GV3101 strain was transformed and infected with onion epidermal cells, and gene expression was observed under confocal microscope. Plant overexpression vector was constructed and infected with tobacco. The change of triterpene content was detected by ultraviolet spectrophotometry. The protein with molecular weight of 62.659 ku was obtained by IPTG induction and SDS-PAGE detection, which was in line with the expected protein, and the expression was the most obvious at 37 鈩，

本文编号：1799828