农杆菌介导油菜油体基因与碱性成纤维细胞生长因子融合基因转化油菜研究

Agrobacterium tumefaciens-mediated Transformation of Brassica campestris L. with Fusion Gene Ycoil-bFGF

[目的]利用植物转基因技术,生产药用蛋白。[方法]以带柄子叶作为转化受体,利用根癌农杆菌介导法将油菜油体基因与bFGF的融合基因转入油菜中,在获得潮霉素(Hyg)抗性植株的同时,对油菜的再生条件进行优化,并对获得的抗性苗进行PCR和Southern检测。[结果]油菜最佳转化再生条件为外植体预培养2 d,共培养3 d,菌液浓度OD值为0.3,侵染时间5 min。[结论]该研究中,融合基因已成功整合到油菜基因组中,为以后转基因植物种子的蛋白提取、分离和纯化奠定了基础。

[ Objective ] This study was to generate medical protein via transgenic plant. [ Method ] Using rapeseed （Brassica campestris L. ） cotyledons with petiole were used as experimental materials, fusion gene of seed oil body-basic fibroblast growth factor（Ycoil-bFGF） was intro- duced into rapeseed via Agrobacterium tumefaciens-mediated method; meanwhile regeneration conditions of rapeseed were also optimized; the regenerated hygmyein resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene has integrated into rapeseed genome; the optimized transformation conditions were as follows ： pre-euhure of explant 2 d, co-cuhure 3 d, bacteria solution OD600 0.3, infection duration 5 min. [ Conclusion] The results laid a solid foundation for extracting, isolating and purifying the protein for transgenic plant.