外源基因Egfp在转基因小鼠中的遗传和表达稳定性研究

The study on inheritance and expression stability of exogenous gene Egfpin transgenic mice

本实验随机抽取不同窝的纯合绿色荧光蛋白(EGFP)转基因小鼠子代进行检测,分别取其尾部组织,提取DNA,质量检测,PCR扩增。经研究发现所有样本在PCR扩增中显示目的条带,对其进行切胶回收纯化,连接转化及克隆测序。测序结果100%匹配且测序峰图的趋势正常。随后对这些转基因小鼠尾组织样本进行定量实验,检测其外源基因Egfp的表达情况,结果发现有一部分小鼠的mRNA表达水平较低。得出的结论是外源基因Egfp稳定地遗传给了后代但在部分后代个体中不能稳定有效表达。本研究通过对外源基因Egfp遗传和表达稳定性的检测,为通过转基因技术培育家畜新品种的研究提供了坚实的理论基础。

In this study,we randomly selected the offsprings of different homozygous enhanced green fluorescent protein （EGFP） transgenic mice to detect：taking the tail tissues,extracting DNA,quality testing and PCR amplification,respectively.We found that all samples in the PCR amplification display target band,then cut agarose gel recycling purified,cloning and sequencing.The sequencing results matched 100％ and the trend of sequencing peak is normal.Subsequently taking the tail tissue samples of transgenic mice for quantitative experiments to detect the expression levels of exogenous gene Egfp,the result showed that a part of transgenic mice had lower levels of mRNA expression.It is concluded that exogenous gene Egfp stably inherited to the offspring,but in some individuals,it couldn＇t be stably and effectively expressed.In our study,by detecting inheritance and expression stability of exogenous gene Egfp provides a solid theoretical foundation on the research about cultivating new varieties of livestock for transgenic technology.