鸡传染性支气管炎病毒S1基因与猪IgG Fc基因在HeLa细胞中的融合表达 |
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引用本文: | 张明富,陈汉阳,彭博,佟铁俦,陈焕春,郭爱珍.鸡传染性支气管炎病毒S1基因与猪IgG Fc基因在HeLa细胞中的融合表达[J].生物技术通报,2008(Z1). |
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作者姓名: | 张明富 陈汉阳 彭博 佟铁俦 陈焕春 郭爱珍 |
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作者单位: | 1. 华中农业大学农业微生物学国家重点实验室,武汉,430070;华中农业大学动物医学院,武汉,430070 2. 华中农业大学农业微生物学国家重点实验室,武汉,430070;华中农业大学动物医学院,武汉,430070;华中农业大学湖北省预防兽医学重点实验室,武汉,430070 |
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基金项目: | 973SARS防制基础研究专项(2003CB514122);;湖北省科技攻关计划(项目编号:2006AA205A02) |
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摘 要: | 根据Genbank中发表的猪IgG Fc段基因及IBV S1基因序列,设计并合成引物。以猪肝组织总RNA为模扩增出猪IgG Fc基因,以含全长IBV M41 S基因的质粒为模板扩增出IBV S1基因,分别克隆至T载体。DNA测序表明,所获得的IBV S1基因大小为1.5 kb,IgG Fc大小为1kb,序列正确。将IBV S1与IgG Fc基因串连,插入含有人组织型纤维蛋白溶酶原激活物分泌信号肽序列(tPA)真核表达载体pcDNA3.1-tPA上,在HeLa细胞上进行瞬时融合表达。经免疫荧光和斑点杂交检测,表达产物同时具有IBV S1蛋白和IgG Fc活性。
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关 键 词: | 禽传染性支气管炎病毒 S1基因 猪IgG Fc基因 真核表达 融合表达 |
The Fusion Expression of Avian Infectious Bronchitis Virus S1 Gene and Porcine IgG Fc Gene in HeLa Cells |
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Abstract: | According to the published gene sequences of IBV M41 S1 gene and porcine IgG heavey chain Fc fragment DNA in GenBank,we synthesized two pairs of specific primers.Total RNA of swine liver was taken as the template to amplify IgG Fc gene by RT-PCR.IBV M41 S1 gene was amplified by PCR from the plasmid encoding full length of S gene. Both of the two genes were respectively cloned into the pMD18T vectors and sequencing were confirmed to be correct by sequencing. The results showed that we have obtained the right... |
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Keywords: | IBV S1 gene Porcine IgG Fc gene Eukaryotic expression Fusion expression |
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