| 稳定表达PRRSV N蛋白的Marc-145细胞系的构建 |
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| 引用本文： | 任同伟,覃念,全东群,谢欣,王豪,王玉旭,陈樱,欧阳康,黄伟坚,韦祖樟.稳定表达PRRSV N蛋白的Marc-145细胞系的构建[J].中国动物传染病学报,2022(1):15-19. |
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| 作者姓名： | 任同伟 覃念 全东群 谢欣 王豪 王玉旭 陈樱 欧阳康 黄伟坚 韦祖樟 |
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| 作者单位： | 广西大学动物科学技术学院动物传染病与分子免疫学实验室 |
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| 基金项目： | 广西自然科学基金(2016GXNSFAA380159,2018GXNSFDA281021);国家自然科学基金(31660716)。 |
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| 摘 要： | 为了构建稳定表达猪繁殖与呼吸综合征病毒(PRRSV)N蛋白的Marc-145细胞系,以PRRSV全长感染性克隆为模板,通过PCR方法扩增PRRSV N基因,将N基因克隆到慢病毒载体中,获得重组质粒pLenti-CMV-N,利用三质粒慢病毒包装系统转染293T细胞,包装成表达N蛋白的慢病毒颗粒,将慢病毒颗粒转导至Marc...
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| 关 键 词： | PRRSV N蛋白 细胞系 慢病毒载体 |
Construction of a Marc-145 Cell Line Stably Expressing PRRSV N Protein |
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| REN Tongwei,QIN Nian,QUAN Dongqun,XIE Xin,WANG Hao,WANG Yuxu,CHEN Ying,OUYANG Kang,HUANG Weijian,WEI Zuzhang.Construction of a Marc-145 Cell Line Stably Expressing PRRSV N Protein[J].Chinese Journal of Animal Infectious Diseases,2022(1):15-19. |
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| Authors: | REN Tongwei QIN Nian QUAN Dongqun XIE Xin WANG Hao WANG Yuxu CHEN Ying OUYANG Kang HUANG Weijian WEI Zuzhang |
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| Institution: | (Laboratory of Animal infectious Diseases and molecular Immunology,College of Animal Science and Technology,Guangxi University,Nanning 530005,China) |
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| Abstract: | In order to construct a Marc-145 cell line stably expressing PRRSV N protein,the N gene was amplifi ed by PCR using an infectious cDNA clone(pGXAM)as template.Then the N gene fragment was cloned into Lentiviral vector,resulting a recombinant clone pLenti-CMV-N.Three-plasmid Lentiviral packaging system containing N gene was then transfected into 293T cells.The packaged lentiviral particles were transducted into Marc-145 cells.The Marc-145 cell lines stably expressing PRRSV N protein were screened by hygromycin B and end-point dilution.Sequencing of PCR products showed the genome of the transducted Marc-145 cell line contained N gene.The expression of the N protein in this cell line was confi rmed by IFA and Western blot.These results showed that a Marc-145 cell line was constructed for stable expression of PRRSV N protein. |
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| Keywords: | PRRSV N protein cell line Lentiviral vector |
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