甜瓜蔗糖合成酶基因(CmSS1)反义表达载体的构建及遗传转化

摘要

以BamH Ⅰ和SalⅠ双酶切pMD18-T-CmSS1和表达载体pBI121,再用T4 DNA连接酶将回收的目的片段反向与pBI121连接.结果证明,CmSS1反向插入到pBI121中,得到了pBI121-CmSS1的重组质粒；利用农杆菌介导法将反义CmSS1基因转化甜瓜,经PCR检测,得到5株转基因植株.这为以后研究甜瓜蔗糖合成酶的活性调节机制及通过基因工程手段研究甜瓜SS的活性奠定了基础.%The pMD18 - T - CmSSl and the expression vector pBI121 were digested respectively with Bam H I and Sal I , and then the target fragments were connected by T4 DNA ligase. The results showed that the CmSSl gene was successfully inserted into the plant expression vector pBI121 and the antisense expression vector pBI121 - CmSSl was obtained. The anti - CmSSl was transformed to muskmelon by Agrobacterium -mediated, and five transgenic plants were obtained by PCR detection. This research laid foundations for further research of regulation mechanisms of muskmelon SS activity by genetic engineering.