中国医科大学学报2012，Vol.41Issue(12)：1068-1072,5.
MultiSite Gateway技术在疟原虫条件性基因打靶载体快速构建中的应用
Application of MultiSite Gateway Technology in High Efficient Construction of Plasmodium Gene-targeting Vector
摘要
Abstract
Objective To construct targeting vector for conditional knockout of ebl gene in Plasmodium yoelii using the Gateway reeombi-natorial cloning system. Methods Three fragments EBL-3U,KBL-5U1 and EBL-5U2+EBL.orf were amplified from Plasmodium yoelii 17xl strain by PCR. Jn order to gel plasmid pCHD-EBL-FRT, multiple DNA fragments were ligated in a defined order and orientation using Mul-tiSite Gateway Technology. To get plasmid PyUIS4/llp,the PyUlS4-5U fragment was amplified by polymerase chain reaction by using genome from Plasmodium yoelii 17x1 strain and replaced the homologous arm of PbUIS4-5U fragment of the plasmid PbUIS4/flp. The plasmids were verified by restriction endonueleases digestion and sequencing. Results The vectors pCHD-EBL-FRT and PyUlS4/flp were correctly con-structed , which was confirmed by enzyme digestion and DNA sequencing. Conclusion The targeting vectors were successfully constructed by using MultiSite Gateway technology. A rapid and high efficient working system based on MultiSite Gateway technology was established for plasmodium gene-targeting vector construction.关键词
疟原虫/质粒/基因打靶/MultiSite Gateway技术Key words
Plasmodium/plasmid/gene targeting/multisite gateway technology分类
医药卫生引用本文复制引用
王各各,李娇娜,杜峰,邓舒,梁佳元,曹雅明,罗恩杰..MultiSite Gateway技术在疟原虫条件性基因打靶载体快速构建中的应用[J].中国医科大学学报,2012,41(12):1068-1072,5.基金项目
国家自然科学基金资助项目(30972572) （30972572）
