| 摘要: |
| 用根癌农杆菌(Agrobacterium tumefaciens)共培养法，将骨形成蛋白-3成熟肽(hBMP-3m)基因导入甘蓝型自交系油菜(Brassica napus L．)品种秦油-3号，并获得了hBMP-3m基因整合到油菜基因组中的植株。试验所用外植体为油菜4～5 d苗龄的下胚轴；根癌农杆菌为LBA4404，其Ti质粒pCAMBIA-hBMP3m含hBMP-3m基因。切取2 mm左右的下胚轴侵染根癌农杆菌后在分化培养基(MS +5.0 mg/L 6-BA +0.3 mg/L NAA)上共培养2d，然后转到附加25 mg/L潮霉素和500 mg L羧苄青霉素的分化培养基上。切下分化出的茎芽，插入附加25 mg/L潮霉素的生根培养基(MS +0.4 mg/L IBA)中，形成完整的植株。经PCR检测和Southern blot分析，证实hBMP-3m基因已插入到油菜细胞基因组中。 |
| 关键词: 人骨形成蛋白-3成熟肽（hBMP-3m） 油菜 下胚轴 根癌农杆菌 |
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| 基金项目:杨陵示范区专项基金，西北农林科技大学生物技术中心开放课题资助项目 |
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| Transformation of Brassica napus with human bone morphogenetic protein 3 mature piptide gene mediated by Agrobacterium tumefaciens |
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| Abstract: |
| Seeds of Brassica napus L.cv."Qinyou-3"were surface-sterilized and germinated hormone-free MS medium.After four to five days,the hypocotyls were excised in such a way.These hypocotyls were used as the explants for tissue culture and genetic transformation.After 2 days of co-cultivation with Agrobacterium tumefaciens strain LBA4404,the plasmid of which contains gene of hBMP-3m,the explants were transferred onto selection medium containing 25 mg/L hygromycin and 500 mg/L carbenicillin.About one month,shoots emerged from the explants inoculated.They were excised and transferred onto rooting medium containing 25 mg/L hygromycin.Whole plants were obtained.Transformation was confirmed by PCR and Southern blot analysis. |
| Key words: Brassica napus hypocotyls human bone morphogenetic protein 3 mature peptide (hBMP-3m) Agrobacterium tumefaciens |
