论文部分内容阅读
为建立外源基因甜菜叶绿体转化体系,利用分子生物学方法构建了包含有编码苏云金芽孢杆菌晶体蛋白基因Btcry1Ac和编码膦丝菌素乙酰转移酶基因bar的甜菜叶绿体转化载体pSKARBt/bar,以甜菜叶绿体基因组中atpB/rbcL做同源片段,以甜菜叶绿体16S启动子和终止子为调控基因,以bar基因为筛选标记基因.基因枪法转化甜菜叶柄,经筛选获得抗性转基因植株.对转基因植株进行外源基因Btcry1Ac和bar的PCR检测、DNA印迹分析,结果表明:外源基因Btcry1Ac和bar确已导入到甜菜叶绿体基因组中.转基因植株除草剂抗性鉴定及其离体叶片虫试鉴定结果表明:转基因植株具有较强的杀虫活性和抗除草剂特性,表达了相应的蛋白质.研究结果还表明:bar基因在植物叶绿体转化中,既可以用作抗性基因,又可用作转化体筛选的标记基因.建立了甜菜叶绿体转化体系.
In order to establish a chloroplast transformation system of exogenous genes, a chloroplast transformation vector pSKARBt / bar containing the crystal protein Btcry1Ac encoding Bacillus thuringiensis and bar encoding phosphinothricin acetyltransferase was constructed by molecular biology method. The genome of atpB / rbcL as a homologous fragment to the sugar beet chloroplast 16S promoter and terminator as a control gene to bar gene selection marker gene by particle gun transformation beet petiole, obtained by screening resistant transgenic plants transgenic plants outside The results showed that the foreign gene Btcry1Ac and bar had indeed been introduced into the chloroplast genome of sugar beet.The resistance of the transgenic plants to herbicide and the identification of their in vitro leaf pests showed that the transgenic The plant has strong insecticidal activity and herbicide resistance and expresses the corresponding protein.The results also show that the bar gene can be used as a marker for transformant selection in plant chloroplast transformation The establishment of a sugar beet chloroplast transformation system.