中国兽医科学2026，Vol.56Issue(1)：50-55,6.DOI:10.16656/j.issn.1673-4696.2026.0008
环形泰勒虫转化细胞表达外源基因系统的构建
Construction of a system for the expressing exogenous genes in the cells transformed by Theileria annulata
摘要
Abstract
To improve the transfection efficiency,this study used TaNM1 cells transformed by thei-leria annulata as the research target,and obtained RNA encoding enhanced green fluorescent protein(eGFP)protein by in vitro transcription(IVT)technology.After capping and purification,eGFP mRNA was transfected into TaNM1 cells by liposomes.Blank cells was set as negative control,while cells trans-fected with p-eGFP plasmid by either liposome or electro-transfection was set as positive control.The expression of eGFP protein in TaNM1 cells was detected by Western-blot,fluorescence microscope and flow cytometry.The results showed that green fluorescence could be observed when eGFP mRNA was trans-fected into TaNM1 cells with liposomes,and the expression of eGFP protein(27 kDa)was also detected by Western-blot,and the transfection efficiency could reach 66.22％by flow cytometry.The p-eGFP plasmid by liposome or electro-transfection could not induce the expression of eGFP in TaNM1 cells,and elec-tro-transfection induced massive cell death.In conclusion,an efficient in vitro transcription system was successfully established,providing a technical support for in-depth research on the infection and pathogenic mechanism of Theileria annulata.关键词
环形泰勒虫/转化细胞/体外转录/脂质体转染Key words
Theileria annulata/transformed cells/in vitro transcription/liposome transfection分类
农业科技引用本文复制引用
刘培琪,赵帅阳,罗金,姚佳丽,殷宏,关贵全..环形泰勒虫转化细胞表达外源基因系统的构建[J].中国兽医科学,2026,56(1):50-55,6.基金项目
甘肃省自然科学基金项目(24JRRA015) （24JRRA015）
国家自然科学基金项目(32202836) （32202836）
国家重点研发计划项目(2023YFD-1802403) （2023YFD-1802403）
甘肃省基础研究创新群体项目(22JR5RA024) （22JR5RA024）
农业科技创新工程项目(CAAS-ASTIP-2016-LVRI) （CAAS-ASTIP-2016-LVRI）
国家肉牛牦牛产业技术体系项目(CARS-37) （CARS-37）
国家寄生虫资源库项目(NPRC-2019-194-30) （NPRC-2019-194-30）
