| 禽网状内皮组织增生症病毒MD-2株感染性克隆的构建与突变分析 |
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| 引用本文： | 黄小洁,孔冬妮,刘丹,陈晓春,杨飞,侯力丹,李俊平.禽网状内皮组织增生症病毒MD-2株感染性克隆的构建与突变分析[J].中国家禽,2020(1):38-43. |
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| 作者姓名： | 黄小洁 孔冬妮 刘丹 陈晓春 杨飞 侯力丹 李俊平 |
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| 作者单位： | 中国兽医药品监察所 |
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| 基金项目： | 国家重点研发计划项目(2016YFD0500809)。 |
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| 摘 要： | 为构建禽网状内皮组织增生症病毒(REV)MD-2株的感染性克隆,将MD-2株的前病毒基因组全长克隆至pMD18-T Simple载体中构建MD-2全长质粒,经转染CEF细胞后进行病毒拯救及鉴定,并对拯救病毒进行了生物学特性研究。结果表明,成功构建了MD-2全长质粒,拯救病毒感染CEF细胞后用间接免疫荧光试验检测到病毒抗原;拯救病毒的生长特性与亲本病毒相比无明显差异。随后对env基因进行了多个位点的定点突变,并拯救出相应的突变株,结果显示env基因第1433位碱基突变后不能拯救出病毒,推断1433位的突变为致死性突变。MD-2株感染性克隆的构建和病毒拯救的研究为继续研究REV基因和蛋白功能提供了技术支持。
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| 关 键 词： | 禽网状内皮组织增生症病毒 感染性克隆 构建 突变 |
Construction and Mutation Analysis of Infectious Clones of Reticuloendotheliosis Virus Strains MD-2 |
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| HUANG Xiaojie,KONG Dongni,LIU Dan,CHEN Xiaochun,YANG Fei,HOU Lidan,LI Junping.Construction and Mutation Analysis of Infectious Clones of Reticuloendotheliosis Virus Strains MD-2[J].China Poultry,2020(1):38-43. |
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| Authors: | HUANG Xiaojie KONG Dongni LIU Dan CHEN Xiaochun YANG Fei HOU Lidan LI Junping |
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| Institution: | (China Institute of Veterinary Drug Control,Beijing 100081) |
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| Abstract: | To construct the infectious clones of reticuloendotheliosis virus(REV) strains MD-2, the REV MD-2 full-length provirus genome was cloned into pMD18-T Simple vetor. The recombinant plasmid was transfected into CEF cells for virus rescue and identification and biological analysis. The results showed that the MD-2 infectious clones was constructed and the virus was rescued from cell transfection successfully. There was no significant difference in the growth characteristics between the rescue virus and the parent virus. The results showed that the virus could not be saved after the 1433 base mutation of env gene. It was concluded that the 1433 mutation was lethal. The construction of MD-2 infectious clone and the study of virus rescue provide technical support for the further study of REV gene and protein function. |
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| Keywords: | reticuloendotheliosis virus infectious clones construction mutation |
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