摘要
以鸡巨噬细胞HD11作为研究对象,分别采用5种不同的脂质体试剂转染和电转方法将含有绿色荧光蛋白(GFP)的质粒转入HD11细胞中,然后通过比较并筛选出适合鸡巨噬细胞HD11的最佳转染方法。结果显示:Lipofectamine 2000 Reagent、Attractene Transfection Reagent和Trans IT-2020 Transfection Reagent几种常见的脂质体转染试剂并不适用于HD11细胞转染。而Coming Transfection Reagent、Lip2000 Transfection Reagent脂质体转染试剂和电转可用于HD11细胞转染;其中,Lip2000 Transfection Reagent转染试剂效果最好。在此基础上,通过进一步优化得出:当DNA(μg)和Lip2000(μL)的比例为1∶1.5时转染效率最高。本研究成功筛选出一种有效的鸡巨噬细胞HD11转染方法,为鸡巨噬细胞的转染及其相关研究提供了实验依据。
In this study, the plasmid containing green fluorescent protein （GFP） was transfected into chicken macrophage cell line HD11 using five different liposome transfection reagents or electroporation method to optimize the transfection method for HD11 cell. The results showed that several liposome transfection reagents including lipofectamine 2000 Reagent, Attractene Transfection Reagent and TransIT - 2020 Transfection Reagent were not suitable for HD11 cell transfection. However, Coming Transfection Reagent, Lip2000 Transfection Rea- gent and electric transduction could be ＇used for transfecting HD11 cells. The best transfection efficiency was observed in HD11 ceils when transiected with Lip2000 Transfection Reagent. Furthermore, the highest transfection efficiency for HD11 cells was obtained when the ratio of plasmid DNA （μg） to Lip2000 （μL） was 1 ： h 5. Therefore, an efficient method for transfection of chicken macrophage HD11 cell line was successfully established and it provided the experimental basis for this cell line transfection and other related investigations.
出处
《畜牧与兽医》
北大核心
2017年第5期72-76,共5页
Animal Husbandry & Veterinary Medicine
基金
国家自然科学基金重大研究计划(91540117)
国家自然科学基金面上项目(81372237)
国家自然科学基金青年基金项目(31602032)
江苏省畜牧学优势学科项目
关键词
鸡巨噬细胞
转染效率
电穿孔法
脂质体法
chicken macrophage HD11
transfection efficiency
electroporation
lipofection
