禽流感病毒表面抗原HA基因转化小麦的初步研究

为了建立利用小麦生产禽流感口服疫苗的方法,选择禽流感病毒H5N1亚型表面抗原HA基因,与谷类作物启动子构建小麦表达载体,经基因枪法转入我国小麦栽培品种郑9023幼胚,在含5 mg/L草铵膦愈伤诱导培养基和分化培养基及含4 mg/L草铵膦生根培养基上筛选后,PCR鉴定获得转HA基因植株,Southern杂交分析转基因T1...

Transformation of wheat with a hemagglutinin gene from H5N1 subtype of avian influenza

To develop a method for the production of avian influenza proteins in wheat as an oral vaccine,a gene encoding hemagglutinin (HA) of avian influenza H5N1 was constructed in a wheat expression vector with promoter of cereal crops. The constructed vector DNA was bombarded into immature embryos of an elite wheat cultivar Zheng 9023. Calluses induced on media supplemented with 5 mg/L phosphinothricin (PPT) were screened and transferred to media containing 4 mg/L PPT for rooting and regeneration. The PPT-resistant green plantlets derived from the rooting media were identified by PCR,and the transgenic wheat plants carrying the HA gene were obtained. Southern blot analysis of T1 transgenic plants demonstrated that the HA gene was indeed integrated into the wheat genome with a single copy integration pattern. These results provide materials for further investigating expression of avian influenza HA gene and accumulation of its protein in wheat seeds.