摘要
【目的】制备转人表皮生长因子(Human epidermal growth factor,hEGF)基因红花植株,为利用植物生物反应器规模化生产hEGF奠定基础。【方法】利用分子生物学方法将植物偏好的双基因hEGF克隆至表达载体pOP上,构建了重组质粒pOP-hEGF-hEGF,采用冻融法将质粒pOP-hEGF-hEGF转入根癌农杆菌EHA105中,通过农杆菌介导方法转化红花,采用PCR检测、Southern blot筛选转基因红花阳性植株。【结果】成功构建了植物特异表达载体pOP-hEGF-hEGF,通过转化红花子叶,共获得了30株转基因红花植株,其中3株鉴定为阳性,转化率为10%;对转化植株进行分子生物学检测,从DNA水平检测结果可知,hEGF基因已整合进红花基因组中。【结论】获得了转hEGF基因的红花株系。
【Objective】Preparation human epidermal growth factor （Human epidermal growth factor,hEGF） gene transgenic safflower plants,aiming to improve large scale production of hEGF from plant bioreactors.【Method】hEGF was cloned to pOP using molecular biology methods and recombinant plasmid pOP-hEGF-hEGF was established.The pOP-hEGF-hEGF was transformed into agrobacterium EHA105 and then transformed to safflower cotyledons by agrobacteriummediated.Positive transgenic plants were determined by PCR and Southern blot analysis.【Result】Plant binary expression vector pOP-EGF was constructed.30 transgenic safflower plants were obtained and 3 positive transgenic plants were identified with a transformation efficiency of 10%.Molecular biology testing showed that hEGF gene was integrated into the safflower genome successfully.【Conclusion】This study obtained safflower plants incorporated with hEGF gene.
出处
《西北农林科技大学学报（自然科学版）》
CSCD
北大核心
2013年第12期162-166,共5页
Journal of Northwest A&F University(Natural Science Edition)
基金
"十二五"国家高新技术研究与发展计划("863"计划)项目(2011AA100606)
教育部博士点基金-青年教师基金项目(20122223120002)
吉林省中医药管理局项目(2012162)
吉林省教育厅项目
吉林农业大学2012年校内博士启动基金项目(201207)
关键词
人表皮生长因子
红花
遗传转化
human epidermal growth factor
safflower
genetic transformation
