黑芥子酶基因pyk10根特异启动子在番茄中的验证

Functional Analysis of Root Specific Promoter of Myrosinase Gene pyk10 in Tomato

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。

Myrosinase are a group of isoenzymes catalyzing the hydrolysis of glucosinolates, pykl0 is a root and hypocotyls specific myrosinase gene from Arabidopsis thaliana. A promoter fragment of 1 454 bp long of pykl0 was cloned by PCR from Arabidopsis thaliana genomie DNA. The plant expression vector of pPykG was constructed by replaced the 35S promote of vector pBinl21 with pykl0 promote fragment. Gus gene as a repoter gene under the control of myrosinase gene（ pyk 10 ）promoter was introduced into Tomato cultivated variety（ Zhong Shu 6 ） via Agrobacterium transformation. A pykl0 promoter fragment was detected by PCR from transgenenic tomato plants. The result of GUS histochemical detection and quantitative analysis showed that the pykl0 promoter could drived gus gene specifically expression in the transgenic tomato roots.