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。
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.
出处
《华中农业大学学报》
CAS
CSCD
北大核心
2013年第1期9-13,共5页
Journal of Huazhong Agricultural University
基金
"863"国家高技术发展专项(2007AA100505)
教育部博士点基金项目(20070504010)资助
关键词
转基因小麦
HIV
单克隆杭体
胚乳
特异表达
transgenic wheat
human immunodefieiency virus
monoclonal antibody
endosperm
specific expression