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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
FullText Text:
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  Data: 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
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  Data: 10.1002/bmb.20430
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– Name: Abstract
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  Data: 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.)
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