本文選題：橡膠草 + HMGR基因��； 參考：《江蘇農(nóng)業(yè)科學(xué)》2017年16期

【摘要】：分析橡膠草HMGR基因的表達(dá)情況,構(gòu)建原核表達(dá)載體,轉(zhuǎn)化BL21菌株并誘導(dǎo)表達(dá)。構(gòu)建細(xì)胞融合表達(dá)載體,轉(zhuǎn)化GV3101菌株并侵染洋蔥表皮細(xì)胞,在共聚焦顯微鏡下觀察基因表達(dá)情況。構(gòu)建植物過表達(dá)載體并侵染煙草,利用紫外分光光度法檢測(cè)三萜含量的變化。通過IPTG誘導(dǎo)和SDS-PAGE檢測(cè),獲得分子量為62.659 1 ku的蛋白,與預(yù)期蛋白相符合,且37℃誘導(dǎo)6 h時(shí)表達(dá)最明顯;通過激光共聚焦顯微鏡觀察,該基因主要在細(xì)胞膜上表達(dá);紫外可見分光光度法表明轉(zhuǎn)基因煙草中三萜的含量高于空白煙草。橡膠草HMGR基因分別在大腸桿菌、洋蔥鱗片內(nèi)表皮和煙草中成功表達(dá)。
[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，

本文編號(hào)：1799828