CMV启动子驱动报告基因EGFP在转基因猪和转基因小鼠不同组织中的表达效率比较  

Comparison of the Expression Levels of EGFP Gene Driven by CMV Promoter in Different Tissues of the Transgenic Pigs(Sus scrofa) and Mice(Mus musculus)

作　　者：杜宝珠[1] 覃玉凤[1] 杨晓亮[1] 孙燕霞[1] 刘志国[1] 丛佩清[1] 陈瑶生[1] 何祖勇[1] 

机构地区：[1]中山大学生命科学学院/有害生物控制与资源利用国家重点实验室,广州510006

出　　处：《农业生物技术学报》2015年第10期1327-1333,共7页Journal of Agricultural Biotechnology

基　　金：国家转基因重大专项(No.2014ZX08006005-005)

摘　　要：CMV启动子是真核表达载体中常用的一种转录调控元件,通常被认为是一种广谱性的强启动子,能够在多种不同类型的细胞中高效启动外源基因的表达,但关于其在动物不同组织中调控外源基因的表达效率的研究相对较少。本研究对CMV-EGFP转基因猪(Sus scrofa)的小肠、肺、胰脏、心脏、肌肉、肾、肝脏、脾脏等组织和转基因小鼠(Mus musculus)的肺、肌肉、肝脏、脑、肾、脾脏等组织进行切片分析,结果发现各组织中EGFP的表达量有较大差异:在转基因猪中,EGFP的表达量在肺脏、小肠和胰腺中最高,在肌肉和心脏次之,在肝脏、脾脏和肾脏中最低;在转基因小鼠中,EGFP的表达量在肺脏中最高,在肝脏、脑和肌肉中次之,在肾脏和脾脏中最低。由此可见,CMV启动子在动物的肺脏中具有较强的启动能力,在肌肉组织中启动能力适中,而在肾脏和脾脏中启动能力较弱。这提示动物不同组织中激活CMV启动子的转录因子的丰度存在较大的差异,因此在建立转基因动物模型时,应根据外源基因的表达部位,考虑是否选用CMV启动子。The CMV promoter is often used as a transcriptional regulatory sequence in eukaryotic expression vectors,and considered as a universal element capable of transcriptional activity in a wide rang of cells in vitro.However,reports documenting the transcriptional activity of this control element in different tissues of transgenic animals are relatively less.In this study,we produced the CMV-EGFP integrated transgenic pigs（Sus scrofa） through somatic cell nuclear transfer technique and CMV-EGFP integrated transgenic mice（Mus musculus） through pronuclear microinjection.Transgenic positive animals were identified through PCR screen.Then the expression level of reporter gene EGFP was investigated in liver,heart and muscle of the transgenic pigs through q PCR assay.The results showed that EGFP is most abundant in heart,and modest in muscle,and relatively less abundant in liver.We further examined the expression pattern of EGFP in a varietyof tissues in both transgenic pigs and mice through detecting the fluorescence signal from cryosections.The results showed that the CMV promoter can drive EGFP expression in all the examined tissues of transgenic pigs and mice,but on closer inspection,we found that the fluorescence intensity of EGFP was obviously different in different tissues,where the signal was most strong in lung,small intestine and pancreas,modest in muscle and heart,and weak in spleen,kidney and liver of transgenic pigs.Similarly,expression level of EGFP was highest in lung,modest in brain,liver and muscle,and lower in spleen and kidney of transgenic mice.These experiment results suggested that the CMV promoter could drive the expression of EGFP in a wide range of tissues,however,its activity varies notably across different tissue,where it is most active in lung,modest active in muscle,and less active in kidney and spleen in both the transgenic pigs and transgenic mice.This study indicates that the abundance of transcription factors capable of activating the CMV promoter varies significantly in different ti

关 键 词：CMV启动子 EGFP 转基因猪 转基因小鼠 

分 类 号：S881.2[农业科学—特种经济动物饲养]