人类免疫缺陷病毒单克隆抗体在小麦胚乳中的表达
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“863” 国家高技术发展专项(2007AA100505)和教育部博士点基金项目(20070504010)资助


Expression of a monoclonal antibody against human immunodeficiency virus in wheat endosperm
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    摘要:

    为了建立利用小麦生产抗人类免疫缺陷病毒( HIV) 单克隆抗体的方法,将植物胚乳特异启动子 AsGlo 分别与该抗体的重链(HC)和轻链(LC)连接构建小麦表达载体,以基因枪法转入小麦品种扬麦 158。转基因 T0~T3代 PCR 鉴定表明,HC 和 LC 在小麦基因组中共整合和稳定遗传。RT-PCR 分析表明,IgG 重链和轻链基因可在小麦胚乳中正常转录。酶联免疫吸附(ELISA)进一步证实了 IgG 在小麦胚乳中的积累和抗原结合活性,表明 IgG 蛋白能够在小麦胚乳中稳定表达,可利用小麦种子生产抗 HIV 抗体。

    Abstract:

    To establish a method of producing a monoclonal antibody against human immunodeficiency virus (HIV) in wheat,two gene sequences coding the heavy chain and the light chain of the antibody were separately constructed in plant expression vectors containing an endosperm-specific promoter from oat and then co-introduced into wheat cultivar Yangmai 158 via particle bombardment.Polymerase chain reaction (PCR) analysis of transgenic wheat plants from T0 to T3 generations showed that the integration and inheritance of the two genes were stable.Reverse transcription-PCR analysis indicated that both genes were normally transcribed in wheat endosperm of transgenic plants.Enzyme-linked immunosorbent assays (ELISA) further confirmed that the antibody produced in transgenic wheat plants had specific antigen-binding capability.These results suggested that mammalian antibody can be stably expressed in wheat endosperm and wheat seeds may be used for the production of antibody with HIV-neutralizing activity.

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齐燕妮,廖玉才,胡祖权,章美云,李和平.人类免疫缺陷病毒单克隆抗体在小麦胚乳中的表达[J].华中农业大学学报,2013,32(1):9-13

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  • 收稿日期:2012-03-31
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