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新城疫病毒HN蛋白和F蛋白结构域共表达对细胞融合的影响

发表时间:2008-02-15  浏览量:1767  下载量:772
全部作者: 陈亚波,陈雪燕,徐程 ,张桂芝,方维焕
作者单位: 浙江大学动物预防医学研究所 浙江省动物预防医学重点实验室;浙江大学动物预防医学研究所 浙江省动物预防医学重点实验室;浙江大学动物预防医学研究所 浙江省动物预防医学重点实验室;浙江大学动物预防医学研究所 浙江省动物预防医学重点实验室;浙江大学动物预防医学研究所 浙江省动物预防医学重点实验室
摘 要: 将重组真核质粒pcDNA3-HN、pcDNA3-HNa、pcDNA3-HNb、pcDNA3-F和pcDNA3-Fa(c)脂质体介导转染HeLa细胞,分别以新城疫病毒阳性血清和制备的相应多抗为一抗,用FITC标记二抗进行间接免疫荧光试验,均可以观察到荧光,说明重组质粒能在HeLa细胞中表达具有免疫反应性的目的蛋白。将pcDNA3-F和pcDNA3-Fa(c)分别与pcDNA3-HN、pcDNA3-HNa和pcDNA3-HNb在脂质体介导下逐一共转染HeLa细胞。转染后24h,Fa(c)片段与HNa、HNb共转染孔就观察到了细胞融合,36h后每个共转染孔都观察到了细胞融合,同时用间接免疫荧光法也检测到即合胞体。单独转染孔均未观察到细胞融合。结果表明:不同组合共转染对细胞融合都有一定促进,其中Fa(c)片段与HNa、HNb共转染促融合更为明显,可能与这三个片段较小易表达且保留了融合所需的结构有关。
关 键 词: 兽医学;新城疫病毒;血凝素—神经氨酸酶;融合蛋白;细胞融合
Title: Cell fusion upon co-expression of structural domains of hemagglutinin-neuraminidase and fusion protein of Newcastle Disease Virus
Author: CHEN Yabo, CHEN Xueyan, XU Cheng, ZHANG Guizhi, FANG Weihuan
Organization: Institute of Preventive Veterinary Medicine, Zhejiang University and Provincial Key Laboratory of Preventive Veterinary Medicine
Abstract: Recombinant plasmids pcDNA3-HN, pcDNA3-HNa, pcDNA3-HNb, pcDNA3-F and pcDNA3-Fa(c) were transfected individually into HeLa cells. Indirect immunofluorescent assay revealed expression of the target proteins in HeLa cell monolayers using NDV positive serum and rabbit polyclonal antibodies. To evaluate the effects of co-expression on cell fusion of HN and F structural domains, pcDNA3-F or pcDNA3-Fa(c) were co-transfected with pcDNA3-HN, pcDNA3-HNa or pcDNA3-HNb respectively into HeLa cells. After culture for 24 hours, cell fusion was observed in the wells co-transfected with pcDNA3-Fa(c) and pcDNA3-HNa, and also those containing pcDNA3-Fa(c) and pcDNA3-HNb. After 36 hours, cell fusion wells of all co-transfected combinations were found and detectable by indirect immunofluorescent assay. However, cell fusion could not be observed in the wells with recombinant plasmid F or Fa(c) alone. These results indicate that co-expression of fragments of HN and F protein can promote cell fusion. We postulate that these fragments contained the structural domains required for cell fusion and were easier to be expressed than the full length proteins.
Key words: veterinary medicine; newcastle disease virus; hemagglutinin-neuraminidase; fusion protein; cell fusion
发表期数: 2008年6月第3期
引用格式: 陈亚波,陈雪燕,徐程 ,等. 新城疫病毒HN蛋白和F蛋白结构域共表达对细胞融合的影响[J]. 中国科技论文在线精品论文,2008,1(3):283-288.
 
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