灵芝-8基因的番茄果实特异性启动子植物表达载体的构建

Construction of LZ-8 expressed vector with its tomato fruit-specific promoter

构建含有灵芝-8(LZ-8)基因和番茄果实特异性E8启动子的重组载体,并将其转化到根瘤农杆菌中。通过PCR法获取LZ-8基因和E8启动子序列,将目的基因和E8启动子序列构建到植物表达载体pB I121中,获得果实特异性表达LZ-8蛋白的重组质粒。并采用PCR、限制性内切酶酶切和序列测定分析法,对重组质粒进行鉴定,将其转入根瘤农杆菌GV3101中。PCR法、限制性内切酶酶切图谱和序列测定分析均表明番茄果实特异性表达LZ-8蛋白的重组质粒构建成功。获得了含有LZ-8基因和E8启动子的重组质粒,并成功转化根瘤农杆菌,为下一步LZ-8蛋白在番茄果实中特异表达奠定基础。

The target gene LZ-8 and the sequence of fruit-specific promoter E8 were acquired by PCR, and pBI121 vector was linked to LZ-8 gene and its promoter ES. The reconstructed plasmid was identified by PCR, restricted enzyme map, and sequence analysis, then was transformed to A. tumefaciens GV3101. The result showed that the reconstructed plasmid carried LZ-8 gene and its promoter E8 by PCR, restricted enzyme map, and sequence analysis. This experiment will build up a foundation for the following study of expressing protein LZ-8 in the tomato fruit.