| 摘要: |
| 【目的】利用基因工程技术获得FGF21转基因油菜植株，为FGF21蛋白生产体系的建立奠定基础。【方法】将人源FGF21基因的cDNA序列，根据植物偏好密码子进行改造，并通过PCR拼接技术扩增FGF21全长基因，将该基因插入油菜油体特异表达载体,通过冻融法转到根瘤农杆菌LBA4404，叶盘法转化甘蓝型油菜“沪油15”，并对转基因油菜进行PCR检测和Southern blot分析，最后对FGF21蛋白在转基因油菜种子中的表达的进行了Western blot分析。【结果】合成了FGF21全长基因，成功构建了含FGF21基因的植物双元表达载体p1390yoFGF21，并建立了油菜遗传转化体系。通过油菜遗传转化获得2株转基因植株，PCR扩增和Southern blot检测证实，重组FGF21基因已整合到油菜基因组中；Western blot分析检测显示，FGF21在转基因油菜种子蛋白中具有一定水平的表达量，并具有良好的抗原性。【结论】FGF21基因被成功整合到油菜基因组中，收获的T0代种子蛋白具有良好的抗原性。 |
| 关键词: 成纤维细胞生长因子-21 甘蓝型油菜 生物反应器 转基因 农杆菌 |
| DOI： |
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| 基金项目:国家“863”高新技术研究与发展计划项目生物反应器重大专项(2007AA100503);吉林省科技发展重点计划项目(20070922);教育部高等学校科技创新工程重大项目(70S018);长春市科技计划项目(06GG150) |
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| Genetic transformation of fibroblast growth factor 21 to Brassica napus mediated by Agrobacterium tumefaciens |
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| Abstract: |
| 【Objective】The study was done to obtain the transgenic plant of FGF21 in rape by genetic technology and establish the foundation for FGF21 expression.【Method】The human fibroblast growth factor 21(FGF21) gene was synthesized by fusion PCR after optimization using plant preferred codons.The recombinant FGF21 was inserted into downstream of oleosin gene of the plant oilbody expression vector,and then was transformed into Brassica napus Huyou 15 by Agrobacterium tumefaciens LBA4404.【Result】 The full gene of FGF21,the plant binary expression vector p1390yoFGF21 and two transgenic plants containing p1390yoFGF21 were successfully obtained.Subsequent PCR and Southern blot confirmed that FGF21 gene was integrated into the genome of oilseed rape.And Western blot also showed that FGF21 in transgenic oilseed rape had a certain amount of expression and good antigenicity.【Conclusion】FGF21 gene was successfully integrated into the genome of rape and gained T0 generation seed,and its protein had good antigenicity. |
| Key words: FGF21 Brassica napus plant bio-reactor transformation gene Agrobacterium tumefaciens |
