禽流感病毒HA基因在拟南芥中的表达  

Expression of a Hemagglutinin Gene Derived from the Avian Influenza Virus H5N1 Strain in Arabidopsis thaliana

作　　者：张鹏[1,2] 廖玉才[3,2] 黄涛[1,2] 瞿波[3,2] 李和平[1,2] 

机构地区：[1]华中农业大学生命科学技术学院,武汉430070 [2]华中农业大学麦类作物分子生物技术实验室,武汉430070 [3]华中农业大学植物科学技术学院,武汉430070

出　　处：《华中农业大学学报》2009年第4期386-389,共4页Journal of Huazhong Agricultural University

基　　金：国家"863"高技术研究发展专项经费(2007AA100505);教育部博士点基金(20070504010)资助

摘　　要：将H5N1亚型禽流感病毒血凝素(HA)基因构建到植物表达载体,经农杆菌法转入拟南芥基因组中,酶联免疫吸附(ELISA)测定分析T2代转基因株系,据OD值筛选获得HA基因高效表达株系;RT-PCR及Western blot分析进一步证实,这些转基因株系均可转录HA基因,翻译形成相应的蛋白质。拟南芥叶片中表达积累的HA蛋白经亲和层析纯化,Western blot分析检测到1条分子量为65 ku的蛋白质带,与预期的HA蛋白大小完全一致,没有发生蛋白质降解现象。表明禽流感病毒HA蛋白可在植物细胞中稳定表达,植物可用于生产禽流感疫苗。A gene coding for hemagglutinin derived from the avian influenza virus H5N1 strain was constructed in a plant expression vector and introduced into Arabidopsis thaliana via Agrobacteriurn-mediated transformation. Enzyme-linked immunosorbent assays of transgenie Arabidopsis T2 lines screened plants with a high level of HA gene expression based on their OD values. Reverse-transcription PCR and Western blot analyses further revealed that the HA gene was properly transcribed and translated into the protein in the selected lines. From the leaves of transgenie Arabidopsis plants, the HA protein was affini- ty-chromatography purified and a distinct protein band, with 65 ku in size, was detected in Western blots. This molecular mass of the HA protein was as expected, with no degradation proteins detectable on the blots. These results suggested that the HA protein from the avian influenza virus can be stably expressed in plant cells and plants could be used for the expression of avian influenza virus proteins potentially used as vaccines.

关 键 词：禽流感病毒 血凝素基因 拟南芥 植物表达蛋白 亲和层析纯化 

分 类 号：Q786[生物学—分子生物学]