黑芥子酶基因pyk10根特异启动子在番茄中的验证.pdfVIP

生物技术通报 ·研究报告· BIOTECHNOLOGY BULLETIN 2012年第11期 黑芥子酶基因 pyk10 根特异启动子在番茄中的验证 崔姗姗  乔亚科  李桂兰  钟磊  刘杰  王林红 （河北科技师范学院植物细胞工程实验室，昌黎 066600） 摘 要 ： 黑芥子酶是一类催化芥子油苷水解的同工酶，pyk10 是一个在拟南芥根和下胚轴中特异表达的黑芥子酶基因。从 拟南芥Columbia 生态型基因组中克隆的长度为1 450 bp 的pyk10 基因启动子片段，以gus 基因为报告基因构建了植物表达载体 pPykG。通过农杆菌介导法，将pyk10 的根特异表达启动子以及gus 基因转入了番茄中蔬6 号，经PCR 检测，转化植株中扩增出 pyk10 启动子特异性条带。组织化学法检测及定量分析，显示pyk10 启动子驱动gus 基因在番茄的根部特定表达。 关键词 ： 拟南芥 pyk10 启动子 gus 基因 番茄 根特异表达 Functional Analysis of Root Specific Promoter of Myrosinase Gene pyk10 in Tomato Cui Shanshan Qiao Yake Li Guilan Zhong Lei Liu Jie Wang Linhong （Plant Cell Engineer Lab ，HeBei Normal University of Science and Technology ，Changli 066600 ） Abstract:  Myrosinase are a group of isoenzymes catalyzing the hydrolysis of glucosinolates. pyk 10 is a root and hypocotyls specific myrosinase gene from Arabidopsis thaliana . A promoter fragment of 1 454 bp long of pyk 10 was cloned by PCR from Arabidopsis thaliana genomic DNA. The plant expression vector of pPykG was constructed by replaced the 35S promote of vector pBin 12 1 with pyk 10 promote fragment. Gus gene as a repoter gene under the control of myrosinase gene（pyk10 ）promoter was introduced into Tomato cultivated variety（Zhong Shu 6 ）via Agrobacterium transformation. A pyk10 promoter fragment was detected by PCR from transgenenic tomato plants. The result of GUS