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斑马鱼胚胎中Cre/lox介导的定点整合
秦瑶; 朱作言; 汪亚平
2007
Source Publication湖北省遗传学会、江西省遗传学会2006年学术年会暨学术讨论会论文摘要集
Abstract为验证Cre重组酶介导的盒式交换反应在斑马鱼中的可行性,本研究采用两种类型的lox突变体和双色荧光报告基因,构建了双交换载体pcMVlox66CreintEGFPlox2272和 Plox71RFPlox2272。将Cre酶基因插入载体pCS2+,获得体外转录模板质粒pCSCre。为检测Cre重组酶对斑马鱼胚胎发育的影响,将体外转录的Cre RNA稀释至不同浓度分别注射入斑马鱼受精卵。结果显示,当Cre RNA浓度高于200ng/μl时会导致高死亡率和高畸形率;而浓度低于 100ng/μl的Cre RNA对胚胎发育不造成影响。进一步将50ng/μl的Cre RNA同双交换载体共注射到斑马鱼受精卵中,筛选到发红色荧光的胚胎。PCR检测和测序鉴定证实,在Cre重组酶作用下,两个载体中lox位点问的荧光基因发生了置换反应。实验结果表明,Cre/lox系统介导的盒式交换反应是实现转基因鱼外源基因定点整合的有效策略。
Language中文
Document Type会议论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/14760
Collection会议论文
Recommended Citation
GB/T 7714
秦瑶,朱作言,汪亚平. 斑马鱼胚胎中Cre/lox介导的定点整合[C],2007.
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