CAJ | 학술논문

采用热激启动子Gmhsp17.5C控制Cre定位重组酶介导的DNA删除系统.在这个系统中,在热激启动子控制下的Cre重组酶的表达导致两侧带有相同方向loxp位点的CaMV35S-GUS片段从转基因烟草(Nicotiana tabacum L.cv.W38)的基因组中删除.通过定量PCR的方法鉴定这个转基因系统,显示了这个系统的重组效率.结果显示在两个小时热激处理后转基因烟草中有41%的CaMV35S-GUS片段被删除.由于热激诱导的定点重组系统有容易操作、对热敏感和无背景表达等优点,因此有利于采用这个系统在转基因植物中进行可诱导的基因操作.
채용열격계동자Gmhsp17.5C공제Cre정위중조매개도적DNA산제계통.재저개계통중,재열격계동자공제하적Cre중조매적표체도치량측대유상동방향loxp위점적CaMV35S-GUS편단종전기인연초(Nicotiana tabacum L.cv.W38)적기인조중산제.통과정량PCR적방법감정저개전기인계통,현시료저개계통적중조효솔.결과현시재량개소시열격처리후전기인연초중유41%적CaMV35S-GUS편단피산제.유우열격유도적정점중조계통유용역조작、대열민감화무배경표체등우점,인차유리우채용저개계통재전기인식물중진행가유도적기인조작.
Cre site-specific recombinase-mediated DNA excision system was driven by the heat shockpromoter Gmhsp17.5C. In this system, the DNA fragment with CaMV35S-GUS franked by two identicalorientation loxp sites could be excised from the transgenic tobacco (Nicotiana tabacum L. cv. W38) by Creexpression under control of heat shock promoter. This transgenic system has been determined by quantitativePCR and showed Cre/lox mediated recombination efficiency. Results showed that 41% of DNA fragmentwith CaMV35S-GUS in the transgenic tobacco could be excised after a two-hour heat shock treatment.Based on several advantages of heat shock-inducible site-specific recombination system such as easymanipulation, sensitivity to heat shock and no background expression, it can be potentially used for induc-ible DNA manipulationin transgenic plant.