González Andrés, Fillat María F
Aragon Institute for Health Research, San Juan Bosco 13, Zaragoza 50009, Spain.
Department of Biochemistry and Molecular & Cell Biology, University of Zaragoza. Pedro Cerbuna 12, Zaragoza 50009, Spain.
Biochem Mol Biol Educ. 2018 Sep;46(5):493-501. doi: 10.1002/bmb.21136. Epub 2018 Aug 1.
Recombinant protein expression and site-directed mutagenesis of target genes have demonstrated an increasing importance in the fields of molecular biology, biochemistry, biotechnology, and medicine. By using the flavodoxin of the model cyanobacterium Anabaena sp. PCC 7120 as a laboratory tool, we designed a comprehensive laboratory practice encompassing several well-established molecular biology techniques and procedures in order to fulfill two main objectives: (1) overexpression and immunodetection of Anabaena flavodoxin in recombinant Escherichia coli cell extracts, and (2) site-directed mutagenesis of the Anabaena flavodoxin gene isiB. This lab practice provides undergraduate students the possibility to perform by themselves several essential techniques in the field. With the aid of professors, students are stimulated to think, to interpret, and to discuss the results based on what they had learned in previous theoretical courses. © 2018 by The International Union of Biochemistry and Molecular Biology, 46(5):493-501, 2018.
重组蛋白表达和靶基因的定点诱变在分子生物学、生物化学、生物技术和医学领域已显示出越来越重要的地位。通过使用模式蓝藻鱼腥藻PCC 7120的黄素氧还蛋白作为实验室工具，我们设计了一个综合实验室实践，涵盖了几种成熟的分子生物学技术和程序，以实现两个主要目标：（1）在重组大肠杆菌细胞提取物中过表达鱼腥藻黄素氧还蛋白并进行免疫检测，以及（2）对鱼腥藻黄素氧还蛋白基因isiB进行定点诱变。这个实验室实践为本科生提供了亲自操作该领域几种基本技术的可能性。在教授的帮助下，激励学生根据他们在之前理论课程中学到的知识来思考、解释和讨论结果。© 2018年，国际生物化学与分子生物学联盟，46(5):493 - 501，2018。
