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当前位置: 首页 > 文章 > 鸭源新城疫病毒F基因在昆虫细胞中的表达和鉴定 中国畜牧兽医 2015,42 (11) 2862-2866
Position: Home > Articles > Expression and Identification of F Gene of Duck Newcastle Disease Virus in Insect Cells China Animal Husbandry & Veterinary Medicine 2015,42 (11) 2862-2866

鸭源新城疫病毒F基因在昆虫细胞中的表达和鉴定

作  者：
王安平;朱善元;王永娟;吴双;左伟勇;洪伟鸣
单  位：
江苏农牧科技职业学院江苏省兽用生物制药高技术研究重点实验室
关键词：
鸭源新城疫病毒;F基因;昆虫细胞;表达
摘  要：
为在昆虫细胞中表达鸭源新城疫病毒(NDV)F蛋白,本试验首先根据鸭源NDV F基因序列设计引物,PCR扩增出F基因,将其克隆至杆状病毒表达载体pFastBac1,获得重组转座载体pFastBac-F,将其转化到大肠杆菌DH10Bac感受态细胞中,经抗性和蓝白斑筛选,获得重组杆状病毒穿梭质粒rBacmid-F,在脂质体介导下转染Sf9昆虫细胞,获得重组杆状病毒rBac-F。Western blotting、间接免疫荧光试验结果均显示表达的重组蛋白能与鸭抗NDV阳性血清发生特异性反应,具有良好的反应原性。结果表明鸭源NDV F蛋白在昆虫细胞中获得了成功表达,为鸭新城疫的预防控制奠定了基础。
译  名：
Expression and Identification of F Gene of Duck Newcastle Disease Virus in Insect Cells
作  者：
WANG An-ping;ZHU Shan-yuan;WANG Yong-juan;WU Shuang;ZUO Wei-yong;HONG Wei-ming;Jiangsu Provincial Key Laboratory of Veterinary Bio-pharmaceutical High Tech Research,Jiangsu Agri-animal Husbandry Vocational College;
关键词：
duck NDV;;Fgene;;insect cells;;expression
摘  要：
In order to express the F protein of duck Newcastle disease virus(NDV)in insect cells,one pair of specific primers was designed according to the published genome sequences of duck NDV to amplify Fgene by PCR,the amplified fragment was cloned into baculovirus expression vector pFastBac1.Then the recombinant vector pFastBac-F was transformed into E.coli DH10 Bac competent cells,and the positive recombinant bacmid rBacmid-F was screened according to the resistance and blue-white plague screening.rBacmid-F was transfected into Sf9 insect cells by liposome,once the cytopathic effect was found,the rBac-F could be aquired.The results of Western blotting and indirect immunofluorescence test showed that the recombinant proteins could be recognized by positive serum,indicating that the protein had good reactiongenicity.These results suggested that F protein of duck NDV had been successfully expressed in insect cells,which laid the foundation for the prevention and control of duck Newcastle disease.

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