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当前位置: 首页 > 文章 > 转乳铁蛋白肽和α干扰素基因的牛胎儿成纤维细胞的制备 畜牧兽医学报 2012,43 (10) 1547-1553
Position: Home > Articles > Construction of Lactoferricin and Interferon-α Genes Transgenic Bovine Fetal Fibroblasts Chinese Journal of Animal and Veterinary Sciences 2012,43 (10) 1547-1553

转乳铁蛋白肽和α干扰素基因的牛胎儿成纤维细胞的制备

作  者：
余大为;张守峰;朱化彬;张锦霞;范宗兴;郝海生;刘岩;赵学明;秦彤;杜卫华
单  位：
中国农业科学院北京畜牧兽医研究所;中国军事医学科学院军事兽医研究所
关键词：
乳铁蛋白肽;α干扰素;转基因;牛胎儿成纤维细胞
摘  要：
旨在构建含有乳铁蛋白肽基因和α干扰素基因的载体,并制备转基因牛胎儿成纤维细胞,为研制抗乳房炎和口蹄疫转基因克隆牛提供供体细胞。本研究构建了由山羊β-酪蛋白启动子启动的牛乳铁蛋白肽基因和由CMV启动子调控的人α干扰素基因的载体,以EGFP基因作为报告基因,neo基因作为筛选基因;分别采用脂质体和电击法转染牛胎儿成纤维细胞,G418筛选得到转基因阳性细胞,并用PCR方法检测转基因细胞中目的基因的整合情况,用Western blotting检测α干扰素蛋白的表达。结果,得到了同时含有牛乳铁蛋白肽基因和人α干扰素基因的转基因牛胎儿成纤维细胞,并且α干扰素蛋白在转基因细胞中能有效表达。
译  名：
Construction of Lactoferricin and Interferon-α Genes Transgenic Bovine Fetal Fibroblasts
作  者：
YU Da-wei1,ZHANG Shou-feng2,ZHU Hua-bin1,ZHANG Jin-xia2,FAN Zong-xing1, HAO Hai-sheng1,LIU Yan1,ZHAO Xue-ming1,QIN Tong1,DU Wei-hua1(1.Institute of Animal Science,Chinese Academy of Agricultural Sciences,Beijing 100193,China; 2.Institute of Military Veterinary,Academy of Military Medical Science,Changchun 130062,China)
关键词：
Lactoferricin;interferon-α;transgene;bovine fetal fibroblasts
摘  要：
The aim of this study was to construct expression vector containing Lactoferricin gene and interferon-α gene which was transfected into the bovine fetal fibroblasts.The transgenic cells got would be used as the donor cells for SCNT to produce the transgenic bovine that would resist mastitis disease and FMD.The vector with a bovine Lactoferrincin B cassette containing a goat β-casein regulatory sequence,a human interferon-α cassette containing the immediate early promoter of CMV,the EGFP gene as reporter and the neo gene as positive selection marker,was constructed.FuGENE Transfection Reagent and Amaxa Nucleofector were used in transfection.After 48 hours,the cells were selected with G418 and detected by PCR.The expression of interferon-α protein was detected by Western blotting.The transgenic bovine fetal fibroblast cell lines were constructed in which Lactoferricin and interferon-α genes were integrated and the expression of interferon-α protein was detected.

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