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

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

[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed （ Brassica campestris L. ） by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows： explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds.

1材料与方法1．1材料1．1．1供试材料。白菜型油菜苏州青，市售。1．1．2菌株及质粒。采用根癌农杆菌LBA4404，该菌株含有植物双元表达载体pCAMBIA1390，油菜油体基因与bFGF的融合基因已整合到该载体上，该融合基因由油菜油体基因启动子启动。植物基因组提取试剂盒购自北京百泰克生物技术有限公司，Southern检测试剂盒为DIG High Prime DNA Labeling and Detection Starter Kit I，购于Roche公司。