人类免疫缺陷病毒单克隆抗体在小麦胚乳中的表达

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

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。

To establish a method of producing a monoclonal antibody against human immunodefi- ciency virus （HIV） in wheat,two gene sequences coding the heavy chain and the light chain of the anti- body 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 inte- gration 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 immu- nosorbent 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 ex- pressed in wheat endosperm and wheat seeds may be used for the production of antibody with HIV-neu- tralizing activity.