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

Expressing profiles of transgenes in different cells and chicken tissues delivered by lentiviral vectors

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。

Cells derived from different species with dissimilar differentiational states including HeLa, 293FT, C127, chicken em- bryo fibroblasts （CEF） and chicken primary germ cells （cPGC） were infected in vitro with recombinant lentiviral vectors which were marked with enhanced green fluorescence proteins gene （EGFP）. Data showed that lentivial vectors could infect all types of cells including normal or cancer cells and primary culture cells or infinite cell lines with higher level of expression. The efficiency of transient expression in HeLa and 293FT was about 1 × 10^8 TU · mL^-1 , 5 × 10^7 TU · mL^-1 in C127, 1 × 10^6 TU · mL^-1 in CEF and 1 × 10^2 TU · mL^-1in cPGC. Transfaction of lentiviral may induce adverse morphological transformation of cells in vitro. Using lentiviral vectors to inject fresh lay eggs of hens, EGFP transgenic chickens （ rate about 53% ） had been got and the signal of green fluorescence was found in different tissues of transgenic chickens with different expressing levels and distributing characters.