摘要
目的:运用Dot-ELISA方法对转基因烟草中所表达的重组hEGF蛋白进行初步检测,以确定是否表达出具有免疫功能的蛋白质。方法:将所合成的hEGF基因置于CAMV35S启动子的驱动下,用农杆菌介导的方法转入烟草中,经过抗性筛选和PCR鉴定出阳性转化子后,提取转化子的总蛋白。总蛋白提取物经透析后,取适当浓度蛋白质点在尼龙膜上,用Dot-ELISA方法对所提取的蛋白质进行测定。结果:结果表明,转基因烟草中的重组蛋白能与抗体发生特异性结合,Dot-ELISA反应后的膜上显示出清晰的棕色斑点。结论:通过实验证明Dot-ELISA方法是一种简便快捷的检测方法,适用于植物基因工程中大规模重组蛋白生产的初步检测。
Objective: To identify the immunity activity of recombinant hEGF with Dot ELISA method in hEGF gene transgenic tobacco plants, Methods: In this study, the synthetic hEGF gene was controlled under CAMV35S promoter and was transformed into tobacco plants via agrobacterium-mediated procedure. Total protein was extracted from the transforms identified by PCR, and extraction was used to test the immunity activity of recombinant hEGF by Dot-ELISA method. Results: According to the result, the synthetic hEGF gene was expressed in transgenic tobacco plants and the recombinant protein could link to the antibody especially. Conclusion: It is an explanation that the Dot-ELISA method is simple and rapid, this method can be used to identify recombinant protein in plant transgenic engineering.
出处
《河南大学学报(医学版)》
CAS
2007年第3期9-12,共4页
Journal of Henan University:Medical Science