不同感受态转化的慢病毒质粒的转染及感染效率的比较  

Comparison of Transfection and Infection Efficiency of Lentiviral Plasmids Transformed by Different Competent Cells

作　　者：才文道力玛 伊敏娜 赵比力格 斯琴 芒来[1,2] 格日乐其木格 Tseendolma;Yiminna;ZHAO Bilig;Siqin;Manglai;Gerelchimeg(College of Animal Science,Inner Mongolia Agricultural University,Hohhot 010018,China;Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation,Hohhot 010018,China)

机构地区：[1]内蒙古农业大学动物科学学院,呼和浩特010018 [2]内蒙古自治区马属动物科学研究与技术创新重点实验室,呼和浩特010018

出　　处：《内蒙古农业大学学报(自然科学版)》2025年第2期1-8,共8页Journal of Inner Mongolia Agricultural University(Natural Science Edition)

基　　金：内蒙古自治区博士研究生科学创新项目(B20231061Z)。

摘　　要：外源质粒转化宿主细胞已成为基因工程和DNA克隆技术的基础与核心,是现代分子生物学的重要技术之一。使用大肠杆菌感受态细胞进行转化试验完成外源基因的表达是实验室常用的手段。但由于大肠杆菌感受态细胞种类繁多,选择基因表达效果理想的感受态细胞是必要的。因此,为获得用于马胚胎成纤维细胞感染慢病毒包装体系的转化感受态,本研究分别用了3种不同的感受态细胞(DH5α、Stbl3和INV110)来转化FUW-tetO-EGFP质粒后,使用293T细胞包装病毒颗粒,之后用收集的病毒颗粒感染293T细胞和马胚胎成纤维细胞,通过观察荧光信号,比较三者的转染及感染效率。结果显示:对于转染效率的比较,使用DH5α感受态细胞转化之后的质粒转染293T细胞的效率显著高于INV110感受态细胞和Stbl3感受态细胞;而对于感染效率的比较,使用Stbl3感受态细胞转化之后的质粒包装的病毒感染293T细胞和马胚胎成纤维细胞的效率都显著高于DH5α感受态细胞和INV110感受态细胞。上述结论表明了Stbl3感受态细胞可用于马胚胎成纤维细胞感染慢病毒质粒转化,这一结果将为建立完整的慢病毒包装体系奠定基础,为后续马的基因功能研究工作提供基础试验数据。The transformation of host cells by exogenous plasmids has become the basis and core of genetic engineering and DNA cloning technology.It is one of the important techniques in modern molecular biology.The use of Escherichia coli receptor cells for transformation experiments to complete the expression of exogenous genes is commonly used in the laboratory.However,due to the wide variety of E.coli receptor cells,it is necessary to select the competent cells with the ideal gene expression effect.Therefore,to obtain the transformed competent cells for the lentiviral packaging system for equine embryonic fibroblast infection,three different competent cells(DH5α,Stbl3,and INV110)were used in this study to transform the FUW-tetO-EGFP plasmid,and then the viral particles were packaged by using 293T cells.Then the collected virus particles were infected with 293T cells and equine embryonic fibroblasts,and the transfection and infection efficiencies of these three were compared by observing the fluorescence signals.The re-sults showed that,in terms of the transfection efficiency,it was found that the plasmid transformed with DH5αwas significantly more effective than both INV110 and Stbl3 in transfecting 293T cells.For the comparison of infection efficiency,the plasmid transformed with Stbl3 was significantly more effective than both DH5αand Stb13 in infecting 293T cells and equine embryonic fibroblasts with the collected viral particles.Additionally,the infection efficiency in equine embryonic fibroblasts was significantly higher than that observed with DH5αand INV110.These results indicated that Stbl3 was capable of transforming the lentiviral plasmid for the infec-tion of equine embryonic fibroblasts,thereby laying the foundation for the establishment of a comprehensive lentiviral packaging sys-tem,and providing the basic experimental data for future research on gene function in horses.

关 键 词：感受态细胞 大肠杆菌 马胚胎成纤维细胞 慢病毒 基因表达 

分 类 号：S821[农业科学—畜牧学] Q81[农业科学—畜牧兽医]