基因组学及应用生物学.pdf

基因组学与应用生物学，2009 年，第28 卷，第2 期，第229-236 页 Genomics and Applied Biology, 2009, Vol.28, No.2, 229-236 研究报告 Research Report 玉米 基因植物表达载体构建及菊花转化 Lc 王娟 韩科厅 戴思兰* 北京林业大学园林学院, 国家花卉工程技术研究中心, 北京, 100083 * 通讯作者, silandai@ 摘 要 Lc 基因是从玉米中分离得到的与花青素合成相关的调节基因，它在多种植物中的异源表达可以增 加花青素的含量。本研究对pBI121 载体Gus基因后的终止子进行2 次PCR 扩增，在原Sac 玉酶切位点后添 加了新的Sac 玉酶切位点，利用组织化学染色法检测，结果表明改造后的载体上的Gus基因能正常表达，终 止子功能正常，载体改造成功。用改造的pBI121N 构建了含有 基因的植物表达载体pBI121N-Lc ，利用农 Lc 杆菌介导法转化菊花叶盘，获得了 19 株生根抗性苗。通过提取抗性苗基因组总DNA，PCR 扩增 基因和 Lc CaMv35S 启动子获得了11 个阳性株系，PCR 结果表明 基因已经转入菊花中。同时，在已获得的转基因植 Lc 株中发现7 个株系的根系有变红的现象。 关键词 玉米, Lc 基因, 载体构建, 菊花, 遗传转化 Construction of Expression Vector and Transformation of Chrysanthemum with Maize Lc Gene Wang Juan Han Keting Dai Silan * College of Landscape Architecture, Beijing Forestry University, National Engineering Research Center for Floriculture, Beijing, 100083 * Corresponding author, silandai@ DOI ：10.3969/gab.028.000229 Abstract Lc gene, a regulatory gene of anthocyanins biosynthesis, is isolated from maize. And the heterologous expression of Lc-mediated protein enhancs the anthocyanins accumulation. The one new restriction enzyme Sac 玉 site was added to terminator behind Gus gene of pBI121 vector by PCR. Gus histochemical assay indicated that Gusgene can express as same as original pBI121 vector and the function of terminator was normal. The expression vector pBI121N-Lc was constructed by Lc gene using reconstruct vector pBI121N. The vector pBI121N-Lc was transferred into riqietaohong ’by -mediated using leaf disk as the ex- Chrysanthemum伊morifolium ‘ Agrobacterium plants and 19 generation resistance plants were obtained. PCR assay shows that 11 of them are positive plants. The