| 摘要: |
| 【目的】应用基因工程技术，获得表达血管内皮生长因子（VEGF）的转基因番茄植株，为以番茄作为生物反应器生产药用蛋白奠定基础。【方法】利用植物偏好的密码子改造合成VEGF基因全长，构建植物表达载体p1390R VEGF，通过农杆菌菌株EHA105介导将其T-DNA区转入到番茄细胞中，再生后获得转基因番茄植株，对其进行分子和蛋白水平检测。【结果】成功构建了植物整株表达载体p1390R-VEGF，建立了高频的番茄再生培养体系；PCR检测、Southern blot和Western blot检测结果表明，VEGF基因已经转入番茄中，并成功得到了表达。【结论】得到了转基因番茄植株和果实，且VEGF蛋白有良好的抗原性。 |
| 关键词: 转基因番茄 血管内皮生长因子 植物生物反应器 农杆菌 |
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| 基金项目:国家高技术研究发展计划（“863”计划）项目（2011AA1000691008)；吉林省科技发展计划重点项目（20070922）；高等学校科技创新工程重大项目培育资金项目（70S018） |
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| Expression of vascular endothelial growth factor（VEGF） in transgenic tomato |
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LI Hongzhi,LIU Jieting,GUO Ran,et al
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Mudanjiang Medical College,et al
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| Abstract: |
| 【Objective】The study was done to obtain the transgenic plant of VEGF in tomato by genetic technology and to establish the foundation for using transgenic tomato to express pharmaceutical proteins.【Method】The VEGF gene was synthesized by the overlap extension PCR method.Then the VEGF gene was put into plant expression vector p1390R,and transformed into the Agrobacterium tumefaciens EHA105 by the freeze-thaw method.Molecule and protein assays were applied on these transgenic tomato lines.【Result】The plant expression vector p1390R-VEGF was construted,and efficient regeneration system was established,DNA PCR and Southern blot results proved that the target gene was successfully integrated into the genome of tomato and Western blot results confirmd the expression of target protein in tomato.【Conclusion】The VEGF gene was successfully expressed in tomato and the transgenic plants and fruits were obtained,and its protein had good antigenicity. |
| Key words: transgenic tomato vascular endothelial growth factor (VEGF) plant bioreactor Agrobacterium tumefaciens |
