【作者】 杨晶； 李海燕； 李校堃；

【Author】 JingYang1 Haiyan-Li1 Xiuming-Lui1 Hongqi-Fu1 Yanfang-Wang1 QingQu2 Xiaokun-Li1 (Jilin Agricultural University,Ministry of Education Bioreactor and Drug Development Research Center,Changchun Jilin China 130118) (2 Jilin University Public Health Colege,Changchun 130021,China)

【机构】 吉林农业大学教育部生物反应器与药物开发工程研究中心； 吉林大学公共卫生学院；

【摘要】 目的:利用红花表达aFGF可以使aFGF的组织创伤修复的活性和红花的恬血通经、散瘀止痛以及跌打损伤等功效累加,直接用于外伤修复;方法:利用分子生物学方法构建aFGF与GFP的融合基因载体,通过农杆菌介导法将其转化到红花中,形成抗性红花愈伤组织后,利用荧光显微镜进行检测;结果:通过PCR分别扩增了aFGF和GFP基因片段,确定出PCR反应体系和最佳反应条件,合成了融合基因片段aFGF-GFP,成功构建了植物荧光表达载体pCAMBIA1390::35S::aFGF-GFP,用其转化红花,获得的抗性愈伤组织在蓝色光激发下能发出强烈的绿色荧光;结论:抗性红花愈伤组织中的GFP表达效果良好,初步判断aFGF基因在植物细胞中也已经表达。更多还原

【Abstract】 Objective:Using safflower as a bioreactor to produce aFGF, which could not only cumulate the activity in tissue wound repair of aFGF and the functions of safflower which conclude activating blood circulation, eliminating stasis to activate blood circulation, relieving the pain, and to heal the injury from fall; this research could also to repair the trauma directly. It could also save the cost of drug isolation and purification, at the same time it could increase the treatment effectiveness of the drugs. Methods:Construction aFGF and GFP fusion gene vector using molecular biology methods,it is transformed into safflowers by Agrobacterium tumefaciens, forming the resistant safflower callus, to detect by fluorescence microscopy; Results:aFGF gene and GFP gene were amplified by PCR reaction。 It is identified PCR reaction system and the best reactions, to acquire aFGF-GFP fusion gene. It was successfully constructed plant fluorescence expression vector pCAMBIA1390::35S::aFGF-GFP, with its transformation safflower, acquired resistance safflower calli . It can send a strong green fluorescence by blue excitation; Conclusion:GFP in resistance safflower calli are good effection, and so aFGF gene in plant cells has also been expressed.更多还原