Bek腺病毒表达载体的构建及其在哺乳类动物细胞中的表达

Construction of adenovirus expression vector of Bek and its expression in mammalian cells

目的构建Bek腺病毒表达载体并观察该载体在成纤维细胞中转基因表达FGFR2-Ⅲc的情况,为研究骨骼发育打下基础。方法利用AdEasy-1腺病毒载体系统构建Bek腺病毒表达载体;该载体感染成纤维细胞后,通过荧光定量PCR(FQ-PCR)检测Bek腺病毒表达载体转基因表达FGFR2-ⅢcmRNA情况。结果Bek腺病毒表达载体感染成纤维细胞后,成纤维细胞有绿色荧光表达,荧光定量PCR检测表明Bek腺病毒表达载体转基因表达FGFR2-ⅢcmRNA效果明显(P<0·05)。结论Bek腺病毒表达载体构建成功,能够转基因表达FGFR2-ⅢcmRNA,为研究骨骼发育和成骨机制提供了基础。

Objective To construct adenovirus expression vector of Bek and observe its expression of FGFR2-me mRNA in fibroblasts for ensuring easy access to the investigation of skeletal development. Methods The adenovirus expression vector of Bek had been constructed by using AdEasy-1 adenovirus vector system. After infecting fibroblasts, the FGFR2-Ⅲ c mRNA which expressed by this vector in fibroblasts was detected by fluorescent quantitive PCR（FQ-PCR）. Results Green fluorescence in the fibroblasts could be observed after they were infected by adenovirus expression vector of Bek. Detection of fluorescent quantitive PCR indicated that the adenovirus expression vector of Bek had a significant effect on transgenically expressing FGFR2-Ⅲc mRNA（P 〈 0. 05）. Conclusion The adenovirus expression vector of Bek has successfully been constructed that can express FGFR2-Ⅲ c by path of transgene