蛋白质免疫印迹技术在大学生创新实验中的应用

在生物功能研究过程中,通常需要从混合蛋白中区分、检测目标靶蛋白,而蛋白免疫印迹技术能较好的实现该目的.蛋白免疲印迹技术是一种先择性地结合被分离混合物中特异蛋白的分析技术,具有灵敏度高、特异性强、分辨率高、分析容量大,可对蛋白进行定性、定量分析,检测蛋白的时空表达变化情况,被广泛应用于蛋白定性实验技术中.为在大学生创新实验中推广该技术,本教学实验以贵州大学本科生为对象,课程的主要内容包括了解蛋白印迹法的原理、目标蛋白载体的构建、蛋白可溶性表达、PAGE胶制作,以及蛋白质免疫印迹试验.该教学可提高学生对蛋白免疫印迹技术原理和实验操作技能的认识和掌握,同时训练学生在科研中的能力.

Application ofWestern Blotting in Innovative Experiment of College Students

In the process of biological function research, it is usually necessary to distinguish and detect the target protein from the mixed protein, and Western blot technology can better achieve this purpose. Protein immunoblotting is a kind of analytical technology that selectively combines the specific proteins in the separated mixture. It has high sensitivity,specificity, high resolution and large analysis capacity. It can be used for qualitative and quantitative analysis of proteins and detect the changes of spatiotemporal expression of proteins. It is widely used in the qualitative experiment of proteins.In order to popularize the technology in the innovative experiment of college students, this teaching experiment is aimed at undergraduates of Guizhou University. The main contents of the course include understanding the principle of western blotting, the construction of target protein vector, the soluble expression of protein, the preparation of page gel, and the protein immunoblot test. This teaching can improve students’ understanding and mastery of Western blotting technology principle and experimental operation skills, and train students’ ability in scientific research.