原核细胞基因表达调控与产物纯化综合实验设计与教学实践  

Comprehensive Experimental Design and Teaching Practice of Gene Expression Regulation and Product Purification in Prokaryotic Cells

作　　者：李欣[1] 赵玉红[1] 赵立青[1] 李小菊[1] 张伟英[1] 石建党[1] LI Xin;ZHAO Yuhong;ZHAO Liqing;LI Xiaoju;ZHANG Weiying;SHI Jiandang(Biological Experimental Center,Colloge of Life Science,Nankai University,Tianjin 300071,China)

机构地区：[1]南开大学生命科学学院生物国家级实验教学示范中心,天津300071

出　　处：《实验室研究与探索》2020年第8期155-159,188,共6页Research and Exploration In Laboratory

基　　金：2019年南开大学自制实验教学仪器设备项目基金(19NKZZYQ);2019年南开大学实验教学课程改革项目基金(2019NKSYJG);南开大学在线课程建设项目基金(NKJG2019013)。

摘　　要：为了提升分子生物学实验教学效果和教学质量,课程组设计了"原核细胞基因表达调控与产物纯化综合实验"。在设定条件下,利用科研常用技术手段对原核细胞目的基因在转录和翻译水平上进行检测,分析启动子利用效率和蛋白表达水平变化情况,并对靶蛋白进行纯化。结果表明,经过1 mmol/L IPTG诱导,原核细胞模式生物大肠杆菌在转录水平上启动子活性明显提高,翻译水平上可见目的蛋白表达量显著增加,并最终通过填料法亲和层析获得良好的蛋白纯化效果。实验通过科学严谨的实验方案以及合理灵活的教学设计,强化了学生对基因表达调控的理解与掌握,拓展了实验教学的广度与深度,进一步推动了分子生物学实验教学的改革和创新。In order to improve the teaching effect and quality of molecular biology experiment teaching,the national biological experiment teaching demonstration center of authors’ school has designed " comprehensive experiment of gene expression regulation and product purification in prokaryotic cells". The experimental content is to use the common techniques of scientific research to detect the transcription and translation of prokaryotic target genes under the set conditions,analyze the utilization efficiency of promoters and the changes of protein expression levels,and purify the target proteins. The results show that the promoter activity of prokaryotic cell model E. coli at the transcription level is significantly increased by 1 mmol/L IPTG,the expression level of the target protein is significantly increased at the translation level,and finally the protein purification effect is obtained by affinity chromatography. Through scientific and rigorous experimental programs and reasonable and flexible teaching design,the experiment can strengthen students’ understanding and mastery of gene expression regulation,expand the breadth and depth of experimental teaching,and further promote the reform and innovation of molecular biology experimental teaching.

关 键 词：基因表达调控 产物纯化 综合实验 教学实践 

分 类 号：G642.0[文化科学—高等教育学]