Lin Shifeng Zhang Shuzhen Yang Xiuhong Chen Qingshan Yang Qingkai Li Wenbin.CONSTRUCTION OF TWO PLANT EXPRESSION VECTORS WITH SENSE ORANTISENSE SOYBEAN RESISTANCE- RELATED GENE SR1[J].Soybean Science,2005,24(02):90-94.[doi:10.11861/j.issn.1000-9841.2005.02.0090]
大豆抗病相关基因 SR1 正反义植物表达载体的构建及遗传转化研究*
- Title:
- CONSTRUCTION OF TWO PLANT EXPRESSION VECTORS WITH SENSE ORANTISENSE SOYBEAN RESISTANCE- RELATED GENE SR1
- Keywords:
- Soybean SR1 gene ; Expression vectors; Transgenic ; Tobacco
- 文献标志码:
- A
- 摘要:
- 实验构建了CaMV35S 启动子控制下的大豆抗病相关基因 SR1 的正反义植物双元表达载体pBISR1(+)和 pBISR1(-)。通过根癌农杆菌叶盘转化法 , 将正义和反义 SR1 基因导入烟草 Ha-vana 425 ,经卡那霉素筛选,获得了抗性植株 。经 PCR和 PCR- Southern 印迹分析 ,证明抗性植株中整和了 SR1 基因, RT - PCR 分析进一步表明正义和反义基因皆能转录为完整的 mRNA ,经疫霉根腐接种及抗病性鉴定表明 ,转反义基因株系和未转基因株系均轻微感病,而转正义基因株系始终没有出现感病症状 。
- Abstract:
- Two plant expression vectors carrying sense or antisense soybean resistance - related gene SR1under the regulation of cauliflower mosaic virus 35s promoter was constructed. Leaf segments of tobaccoHavana 425 were infected by Agrobacterium tumefaciens LBA4404 with pBISR1(+)or pBISR1(-), fromwhich kanamycin resistant plants were obtained. PCR and PCR - Southern analysis proved that the SR1gene was integrated into the genomes of the tobacco plants, and RT - PCR analysis proved that sense orantisense gene was transcripted into a complete mRNA. The disease resistance assay showed that plantswith antisense gene and control plants were slightly susceptible , and plants with sense gene were not sus-ceptible.
参考文献/References:
1 杨秀红. 大豆抗病相关基因的克隆研究[ D] . 东北农业大学, 农学博士学位论文:2002.
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备注/Memo
?基金项目:863 计划(2003AA207060- 4)资助项目