登革病毒转基因载体pB［PUBnls_EGFP_prM］的构建及其在C6/36细胞中整合作用的检测

利用转基因技术来探索新的蚊媒疾病防治方法，将登革病毒前膜蛋白基因prM重组入以转座子piggyBac因子为基础的载体，构建了昆虫转基因载体pB［PUBnls-EGFP-prM］，在辅助质粒的作用下共同转染白纹伊蚊Aedes albopictus C6/36细胞。PCR和Southern blot证明构建的转基因载体可以将EGFP-prM基因整合入蚊虫基因组中。验证了转座子piggyBac因子、启动子polyubiquitin可以在白纹伊蚊中发挥功能,为进一步构建不传播登革病毒的转基因白纹伊蚊奠定了基础。

Construction of transgenic vector pB [PUBnls-EGFP-prM] and identification of its integrational function in mosquito cell

To explore a new method to control arthropod-borne diseases, prM gene from dengue virus was inserted into a transposon piggyBac based plasmid and an insect transgenic vector pBPUBnls-EGFP-prM] was constructed. The vector was used to transfect Aedes albopictus C6/36 cell with a helper plasmid. PCR and Southern blot tests showed that EGFP-prM gene had integrated into C6/36 cell genome. The results indicate that transposon piggyBac element is functional in Ae. albopictus and should be useful in further constructing dengue-refractory transgenic Ae. albopictus.