过表达紫杉醇合成关键酶基因是提高红豆杉细胞紫杉醇产量的有效方法.本文通过构建C-l3苯丙素侧链CoA-乙酰转移酶(C-13 phenylpropanoid side chain-CoA acetyltransferase,BAPT)基因Bapt的表达载体p1303-SbaptN,采用根癌农杆菌介导法转...过表达紫杉醇合成关键酶基因是提高红豆杉细胞紫杉醇产量的有效方法.本文通过构建C-l3苯丙素侧链CoA-乙酰转移酶(C-13 phenylpropanoid side chain-CoA acetyltransferase,BAPT)基因Bapt的表达载体p1303-SbaptN,采用根癌农杆菌介导法转化中国红豆杉细胞,经潮霉素抗性筛选获得转基因细胞系.转基因细胞的PCR和Southern印迹分析结果表明,T-DNA及其包含的基因与宿主细胞染色体成功整合;Western印迹结果表明,转基因细胞中报告基因GusA-mgfp5正常表达;QRT-PCR结果显示,转基因细胞的Bapt mRNA表达量是未转化细胞的1.26倍;HPLC检测结果显示,转基因细胞的紫杉醇产量为37.4μg/g,是未转化细胞的1.87倍.本研究结果表明,过表达Bapt基因提高了中国红豆杉细胞的紫杉醇产量.展开更多
In plant, evocation of secondary metabolism is associated with complex biochemical and molecular events that are regulated by developmental and environmental factors. In order to get more information about Taxol biosy...In plant, evocation of secondary metabolism is associated with complex biochemical and molecular events that are regulated by developmental and environmental factors. In order to get more information about Taxol biosynthesis, comparison of mRNA populations from Taxus chinensis cells during Taxol-synthesis phase and those during non-Taxol-synthesis phase were performed by mRNA differential display. The results suggested that genes specifically expressed in the Taxol-synthesis phase might be involved in Taxol biosynthesis.展开更多
文摘In plant, evocation of secondary metabolism is associated with complex biochemical and molecular events that are regulated by developmental and environmental factors. In order to get more information about Taxol biosynthesis, comparison of mRNA populations from Taxus chinensis cells during Taxol-synthesis phase and those during non-Taxol-synthesis phase were performed by mRNA differential display. The results suggested that genes specifically expressed in the Taxol-synthesis phase might be involved in Taxol biosynthesis.
