论文部分内容阅读
目的:运用PAdEasy系统构建了带荧光蛋白的人CTLA-4Ig腺病毒载体,为进一步研究CTLA-4Ig诱导免疫耐受的机制奠定了基础。方法:采用基因技术,将目的基因克隆至穿梭质粒pAdTrack-CMV中,利用PAdEasy系统进行重组,然后在293细胞中进行扩增。并进行病毒滴度测定。体外感染人肾小管上皮细胞。结果:酶切鉴定和PCR证明重组人CTLA-4Ig腺病毒载体构建正确。结论:应用PAdEasy细菌内重组技术成功构建了人CTLA-4Ig带荧光腺病毒载体。
OBJECTIVE: To construct human CTLA-4Ig adenovirus vector with fluorescent protein by using PAdEasy system, which lays the foundation for further study on the mechanism of CTLA-4Ig induced immune tolerance. Methods: The gene was cloned into the shuttle plasmid pAdTrack-CMV by gene technology and recombined by PAdEasy system and then amplified in 293 cells. And the virus titer was measured. In Vitro Infection of Human Renal Tubular Epithelial Cells. Results: Restriction endonuclease digestion and PCR proved that recombinant human CTLA-4Ig adenovirus vector was constructed correctly. Conclusion: Human CTLA-4Ig fluorescent adenovirus vector was successfully constructed by PAdEasy bacterial recombination technique.