八氢番茄红素脱氢酶(Pds)基因果实特异表达载体的构建

Construction of Phytoene Desaturase (Pds) Gene Expression Vector with Tomato Fruit Specific E8 Promoter

八氢番茄红素脱氢酶(Pds)是番茄红素合成的关键酶,本研究通过PCR法获取pds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pMD1中,构建了含果实特异表达启动子的pds基因的植物表达载体。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。结果表明:番茄果实特异性表达pds的重组质粒构建成功;通过农杆菌直接转化技术将其成功转入转化农杆菌LBA4404、EHA105中,为下一步pds在番茄果实中特异表达奠定了基础。

Phytoene desaturase （Pds） is a key enzyme in the biosynthesis pathway of carotenoid. The target gene pds and the sequence of tomato fruit specific E8 promoter were amplified by PCR and then cloned into pMD1 vector, respectively. PCR, restriction enzyme map, and sequence analysis were used to identify the reconstructed plasmid. The experimental results confirmed the successful construction of a recombinant plasmid carrying pds gene and promoter E8. The recombinant plasmid successfully transformed Atumefaciens LBA4404 and EHA105. These investigations will provide references for further study of pds expression in tomato fruits.