A Practical Teaching Course in Directed Protein Evolution Using the Green Fluorescent Protein as a Model
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| Title: | A Practical Teaching Course in Directed Protein Evolution Using the Green Fluorescent Protein as a Model |
|---|---|
| Language: | English |
| Authors: | Ruller, Roberto, Silva-Rocha, Rafael, Silva, Artur, Schneider, Maria Paula Cruz, Ward, Richard John |
| Source: | Biochemistry and Molecular Biology Education. Jan-Feb 2011 39(1):21-27. |
| Availability: | John Wiley & Sons, Inc. Subscription Department, 111 River Street, Hoboken, NJ 07030-5774. Tel: 800-825-7550; Tel: 201-748-6645; Fax: 201-748-6021; e-mail: subinfo@wiley.com; Web site: https://secure.interscience.wiley.com/cgi-bin/jhome/112782101 |
| Peer Reviewed: | Y |
| Page Count: | 7 |
| Publication Date: | 2011 |
| Intended Audience: | Teachers |
| Document Type: | Journal Articles Reports - Descriptive |
| Education Level: | Higher Education |
| Descriptors: | Chemistry, Biotechnology, Engineering, Scientific Research, Scientific Concepts, Models, Teacher Education, Foreign Countries, Graduate Students, Science Education, Higher Education |
| Geographic Terms: | Brazil |
| DOI: | 10.1002/bmb.20430 |
| ISSN: | 1470-8175 |
| Abstract: | Protein engineering is a powerful tool, which correlates protein structure with specific functions, both in applied biotechnology and in basic research. Here, we present a practical teaching course for engineering the green fluorescent protein (GFP) from "Aequorea victoria" by a random mutagenesis strategy using error-prone polymerase chain reaction. Screening of bacterial colonies transformed with random mutant libraries identified GFP variants with increased fluorescence yields. Mapping the three-dimensional structure of these mutants demonstrated how alterations in structural features such as the environment around the fluorophore and properties of the protein surface can influence functional properties such as the intensity of fluorescence and protein solubility. (Contains 3 figures, 1 table, and 1 footnote.) |
| Abstractor: | As Provided |
| Number of References: | 16 |
| Entry Date: | 2011 |
| Accession Number: | EJ916138 |
| Database: | ERIC |
| Abstract: | Protein engineering is a powerful tool, which correlates protein structure with specific functions, both in applied biotechnology and in basic research. Here, we present a practical teaching course for engineering the green fluorescent protein (GFP) from "Aequorea victoria" by a random mutagenesis strategy using error-prone polymerase chain reaction. Screening of bacterial colonies transformed with random mutant libraries identified GFP variants with increased fluorescence yields. Mapping the three-dimensional structure of these mutants demonstrated how alterations in structural features such as the environment around the fluorophore and properties of the protein surface can influence functional properties such as the intensity of fluorescence and protein solubility. (Contains 3 figures, 1 table, and 1 footnote.) |
|---|---|
| ISSN: | 1470-8175 |
| DOI: | 10.1002/bmb.20430 |
