以生菜、洋葱、萝卜为试材,用转人植物表达载体pSH-NGN的根癌农杆菌EHA105分别采用真空渗透法和直接注射法瞬时表达ChIFN-γ蛋白,研究其最佳宿主植物和最佳方法。结果显示,真空渗透法和直接注射法在瞬时表达效率上差异不大,真空渗透操作上更简便、更易掌握;新鲜市售生菜,真空抽气25min,共培养3d,瞬时表达效率达到3645pg/g,ChIFN-γ 蛋白得到高效表达。
an experiment was conducted to investigate the transient expression mediated by agrobaeterium using the leaves of lettuce (lactuca sativa L.sp), onion (Allium cepa) and radish (Raphanus sativus L.) as host plants and the agrobacterium tumefaciens EHA 105 containing the plant expression vector pSH- NGN by vacuum method and injection method respectively for genetic transformation and detect ChIFN-γ protein expression index to determine the optimal transformation condition. Comparison with injection method, vacuum method was easy and simple to handle, and had the same transient expressing efficiency. When fresh merchant lettuce acted as host, a higher ChIFN - γ transient expression level of 3 645 pg/g could be obtained as vacuum ejection of 25 min and co-culture for 3 days.