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  

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

作　　者：葛春喜[1] 黄炯烈[1] 陈观今[1] 吴瑜[1] 于洪枫[1] 

机构地区：[1]中山大学中山医学院病原生物学教研室,广州510080

出　　处：《昆虫学报》2003年第5期549-552,共4页Acta Entomologica Sinica

基　　金："2 11工程"重点学科建设基金资助项目 ( 9812 4)

摘　　要：为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AedesalbopictusC6 36细胞。PCR和Southernblot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 。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 pB[PUBnls-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.

关 键 词：白纹伊蚊C6/36细胞 prM基因 piggyBac因子 转座子 

分 类 号：Q966[生物学—昆虫学]