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慢病毒载体介导外源基因在不同细胞及鸡体不同组织中的表达

DOI: 10.7685/j.issn.1000-2030.2008.03.019, PP. 97-101

Keywords: 慢病毒载体,鸡原生殖细胞(cPGC),转基因鸡,绿色荧光蛋白

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Abstract:

以增强型绿色荧光蛋白(EGFP)为标记基因构建复制缺陷型慢病毒载体生产病毒,以病毒感染不同来源、不同分化特征的细胞,并进行鸡囊胚注射,观测外源基因的表达情况。试验结果表明,在所有类型细胞中均可检测到EGFP较高水平的表达,其中在HeLa、293FT细胞中的转染效率约1×108TU.mL-1(TU转染单位),在C127细胞中为5×107TU.mL-1,在鸡胚胎成纤维细胞(CEF)中约为1×106TU.mL-1,病毒在鸡原生殖细胞(cPGC)中转染效率最低为1×102TU.mL-1。病毒的感染可引起细胞表型的不利改变。用该病毒载体注射鸡囊胚获得了可直接活体观察绿色荧光信号的转基因鸡,转基因效率约53%;外源基因在鸡体不同组织如表皮、消化道等器官中检测到不同水平的表达,并表现出不同的分布特征。

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