条件性定点基因捕获载体的构建

基因捕获是依赖于捕获载体的随机插入产生突变而进行基因功能研究的一类重要技术。由于传统捕获载体在捕获位点上的局限性，各种新型的捕获载体正在不断地被设计并应用于基因捕获技术。以PL-451为基础质粒，结合定点的重组酶识别位点LoxP及Frt序列，突变LoxP的1个碱基的LoxP511A和LoxP511B，SA位点以及不含启动子但有poly(A)的EGFP片断，构建了能够在捕获元件两侧克隆同源臂的条件性基因捕获载体，从而为实现可调控性基因捕获奠定基础。

Construction of Conditional Target-trapping Vector

Gene trapping is a powerful method for gene analysis relying on mutation generated by random insertion. Various of new kinds trapping vectors have been endlessly designed and applied for trapping because of the local of conventional trapping vector. Based on plasmid PL-451,combinding with the site specific recombination target sites—Loxp,Frt and LoxP511A,LoxP511B which mutant one base of the LoxP,we constructed a conditional gene trapping vector.The trapping vector contains multicloning sites for cloning of the homology regions,which make it can realize the target trapping depending on homologous recombination. Furthermore, the exprssion of promoterless EGFP with splice acceptor（SA）and polyA in the vector can tagging the work of the trapping cassettes.Construction of this vector establish substantial foundation for conditional trapping of target gene.