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含CpG序列PRRSV SD2 E基因真核表达载体质粒构建及表达 被引量：1

Construction and Expression of Eukaryotic Expression Vector Containing PRRSV SD2 E Gene and CpG Motifs
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摘要 为了提高猪生殖与呼吸综合征病毒山东株(PRRSV SD2株)基因免疫的效果,将PRRSV SD2 E基因插入哺乳动物表达载体PVAX1中,构建出PVAX1-E质粒。再将已筛选出的CpG-ODN序列通过在其两端的粘性酶切末端定向插入含PVAX 1-E的真核表达载体,构建含CpG基因序列真核重组表达质粒CpG-pVAX 1-E。将重组质粒转染COS-7细胞,经RT-PCR检测E基因mRNA的转录和间接免疫荧光试验(IFA)证实:CpG-pVAX 1-E可表达PRRSV GP5蛋白。试验结果为进一步研究PRRSV SD2 ORF5动物免疫反应奠定基础。 To improve the effect of the gene immunization against porcine reproductive and respiratory syndrome virus（PRRSV）SD2 we constructed the eukaryotic expression vector CpG-pVAX1-E containing E gene coding region and CpG motif by cloning E gene segment with CpG motif into eukaryotic expression pVAX1 vector. After conformed by identified by enzyme analysis and nucleotide sequencing test , the recombinant expression vector plasmid CpG-pVAX1-E was transfected into COS-7 ：cells by using lipofectamine methods. The ORF5 mRNA of transfected cells were detected by RT-PCR. The transient expression of PRRSV SD20RF5 nucleocapsid protein was detected by indirect immunofluorescence assay（IFA）. In some transfected COS-7 cells, the green fluorescence was showed, thus we can conclude that the eukaryotic expression vector CpG-pVAX1-E was successfully constructed and expressed in vitro, which will lay a foundation for studying on further immunogenicity of PRRSV SD2 ORF5 DNA vaccine.
作者 温建新 李俊 周顺 任慧英 刘文华 邹玲 徐文 牛钟相 王金宝
机构地区 山东农业大学动物科技学院 山东省农业科学院 青岛农业大学动物科技学院
出处 《西北农业学报》 CSCD 北大核心 2008年第3期1-6,共6页 Acta Agriculturae Boreali-occidentalis Sinica
基金 山东省自然科学基金项目资助(编号:Z2003D01)
关键词 猪生殖与呼吸综合征病毒山东SD2株 E基因 真核表达载体 瞬时表达 Porcine reproductive and respiratory syndrome virus（PRRSV）SD2 Eukaryotic expression vector E Gene Transient expression
分类号 S852.65 [农业科学—基础兽医学] Q78 [生物学—分子生物学]
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共引文献938

同被引文献14

  • 1温建新,李俊,周顺,任慧英,刘文华,邹玲,徐文,牛钟相,王金宝.不同序列的CpG-ODN对猪体外免疫细胞免疫刺激作用的研究[J].西北农业学报,2007,16(4):71-75. 被引量：10
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西北农业学报
2008年 第3期

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