腺病毒载体转染水牛不同原代体细胞的效果比较  

Effect Comparison of Adenovirus Vector Transfer into Different Buffalo Cells

作　　者：段安琴 庞春英[1] 朱鹏[1] 陆杏蓉 邓廷贤[1] 梁贤威[1] 

机构地区：[1]中国农业科学院广西水牛研究所,水牛遗传繁育重点实验室,南宁530001

出　　处：《中国畜牧兽医》2016年第10期2661-2665,共5页China Animal Husbandry & Veterinary Medicine

基　　金：农业部转基因重点项目(2014ZX0801012B);广西自然科学基金重点项目(2014GXNSFDA118017);广西科技攻关国际交流与合作项目(桂科合14123001-5);广西水牛研究所基本业务费(1504005)

摘　　要：试验旨在比较腺病毒载体转染水牛乳腺上皮细胞、卵丘细胞、成纤维细胞的效率。将腺病毒质粒pBHGloxdelE13cre、pDC316-eGFP脂质体转染293细胞,收集病毒并测定病毒滴度。用腺病毒载体按不同的感染复数(MOI)感染水牛乳腺上皮细胞、卵丘细胞、成纤维细胞,72h后倒置荧光显微镜下观察,统计感染效率。MOI为25、50、100、200、400时,成纤维细胞感染效率分别为0.7%、7.0%、9.0%、12.5%、34.0%,卵丘细胞感染效率分别为42.5%、55.3%、57.4%、76.0%、80.0%,乳腺上皮细胞感染效率分别为88.7%、100%、100%、100%、100%。结果表明,腺病毒转染乳腺上皮效率最佳,卵丘细胞次之,成纤维细胞效果较差。Buffalo mammary epithelial cell,cumulus cell and fibroblasts were transfected by adenovirus vectors and compared their transfection efficiency.293 cells were transfected with pBHGloxdelE13 cre and pDC316-eGFP by liposome,the virus was collected and titer was detected.Buffalo mammary epithelial cell,cumulus cell and fibroblasts were exposed to different multiplicity of infection（MOI）of adenovirus vectors.After 72 h,the cells were observed with inverted fluorescence microscope,and transfection efficiency was calculated.When the MOI was 25,50,100,200 and 400,the transfection efficiency of fibroblasts were 0.7%,7.0%,9.0%,12.5% and 34.0%,the transfection efficiency of cumulus cells were 42.5%,55.3%,57.4%,76.0% and 80.0%,the transfection efficiency of mammary epithelial cells were 88.7%,100%,100%,100% and 100%.The results showed that the transfection efficiency of mammary epithelial cell was the best,followed by cumulus cell,and fibroblast was poor.

关 键 词：水牛 腺病毒载体 乳腺上皮细胞 卵丘细胞 成纤维细胞 

分 类 号：S852.6[农业科学—基础兽医学]