{"title":"草甘膦氧化酶定向进化及检测草甘膦的化学发光系统:生物技术综合实验室实践实验","authors":"Chanjuan Li, Gaobing Wu, Xuezhu Fan, Siqi Guo, Yuqing Qin, Yonggang Hu, Lifang Ruan","doi":"10.1002/bmb.21721","DOIUrl":null,"url":null,"abstract":"<p>This article describes a comprehensive practical laboratory method for developing an enzyme to more easily measure glyphosate levels in solution. Through this article, undergraduate students of biology majors can conduct research experiments in critical fields by utilizing various techniques, such as chemiluminescence (CL) biosensors with engineered enzymes and are guided in molecular biology laboratories. A glyphosate oxidase mutant library was constructed by DNA shuffling, and a glyphosate oxidase variant with increased glyphosate degradation activity was selected by using a high-throughput screening assay. Following protein overexpression in <i>Escherichia coli</i> (DE3) and purification by affinity chromatography, the glyphosate oxidase variant protein combined with luminol–H<sub>2</sub>O<sub>2</sub> reaction was constructed as a new CL biosensor for detecting glyphosate in soils.</p>","PeriodicalId":8830,"journal":{"name":"Biochemistry and Molecular Biology Education","volume":"51 3","pages":"302-311"},"PeriodicalIF":1.2000,"publicationDate":"2023-03-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Directed evolution of glyphosate oxidase and a chemiluminescence system for glyphosate detection: A comprehensive practical laboratory experiment on biotechnology\",\"authors\":\"Chanjuan Li, Gaobing Wu, Xuezhu Fan, Siqi Guo, Yuqing Qin, Yonggang Hu, Lifang Ruan\",\"doi\":\"10.1002/bmb.21721\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>This article describes a comprehensive practical laboratory method for developing an enzyme to more easily measure glyphosate levels in solution. Through this article, undergraduate students of biology majors can conduct research experiments in critical fields by utilizing various techniques, such as chemiluminescence (CL) biosensors with engineered enzymes and are guided in molecular biology laboratories. A glyphosate oxidase mutant library was constructed by DNA shuffling, and a glyphosate oxidase variant with increased glyphosate degradation activity was selected by using a high-throughput screening assay. Following protein overexpression in <i>Escherichia coli</i> (DE3) and purification by affinity chromatography, the glyphosate oxidase variant protein combined with luminol–H<sub>2</sub>O<sub>2</sub> reaction was constructed as a new CL biosensor for detecting glyphosate in soils.</p>\",\"PeriodicalId\":8830,\"journal\":{\"name\":\"Biochemistry and Molecular Biology Education\",\"volume\":\"51 3\",\"pages\":\"302-311\"},\"PeriodicalIF\":1.2000,\"publicationDate\":\"2023-03-27\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Biochemistry and Molecular Biology Education\",\"FirstCategoryId\":\"95\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/bmb.21721\",\"RegionNum\":4,\"RegionCategory\":\"教育学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"BIOCHEMISTRY & MOLECULAR BIOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochemistry and Molecular Biology Education","FirstCategoryId":"95","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/bmb.21721","RegionNum":4,"RegionCategory":"教育学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
Directed evolution of glyphosate oxidase and a chemiluminescence system for glyphosate detection: A comprehensive practical laboratory experiment on biotechnology
This article describes a comprehensive practical laboratory method for developing an enzyme to more easily measure glyphosate levels in solution. Through this article, undergraduate students of biology majors can conduct research experiments in critical fields by utilizing various techniques, such as chemiluminescence (CL) biosensors with engineered enzymes and are guided in molecular biology laboratories. A glyphosate oxidase mutant library was constructed by DNA shuffling, and a glyphosate oxidase variant with increased glyphosate degradation activity was selected by using a high-throughput screening assay. Following protein overexpression in Escherichia coli (DE3) and purification by affinity chromatography, the glyphosate oxidase variant protein combined with luminol–H2O2 reaction was constructed as a new CL biosensor for detecting glyphosate in soils.
期刊介绍：
The aim of BAMBED is to enhance teacher preparation and student learning in Biochemistry, Molecular Biology, and related sciences such as Biophysics and Cell Biology, by promoting the world-wide dissemination of educational materials. BAMBED seeks and communicates articles on many topics, including:
Innovative techniques in teaching and learning.
New pedagogical approaches.
Research in biochemistry and molecular biology education.
Reviews on emerging areas of Biochemistry and Molecular Biology to provide background for the preparation of lectures, seminars, student presentations, dissertations, etc.
Historical Reviews describing "Paths to Discovery".
Novel and proven laboratory experiments that have both skill-building and discovery-based characteristics.
Reviews of relevant textbooks, software, and websites.
Descriptions of software for educational use.
Descriptions of multimedia materials such as tutorials on various aspects of biochemistry and molecular biology.
京公网安备 11010802042870号
京ICP备2023020795号-1
分享
求助内容：
应助结果提醒方式：
扫码关注我们
