In Vitro synthesis and activity of reporter proteins in an Escherichia coli S30 extract system: An undergraduate experiment

This undergraduate laboratory experiment integrates multiple techniques (in vitro synthesis, enzyme assays, Western blotting) to determine the production and detection sensitivity of two common reporter proteins (β‐galactosidase and luciferase) within an Escherichia coli S30 transcription/translatio...

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Bibliographic Details
Published inBiochemistry and molecular biology education Vol. 33; no. 6; pp. 426 - 430
Main Author Higgins, Pamela J.
Format Journal Article
LanguageEnglish
Published USA John Wiley & Sons Inc 01.11.2005
John Wiley & Sons, Inc
Subjects
Biochemistry
Data Analysis
In vitro synthesis
Laboratory Experiments
luciferase
Microbiology
reporter protein
Research Design
Science Experiments
Science Instruction
Teaching Methods
Undergraduate Students
β‐galactosidase
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Summary:This undergraduate laboratory experiment integrates multiple techniques (in vitro synthesis, enzyme assays, Western blotting) to determine the production and detection sensitivity of two common reporter proteins (β‐galactosidase and luciferase) within an Escherichia coli S30 transcription/translation extract. Comparison of the data suggests that luciferase is the more suitable reporter for this specific in vitro extract system. Simple modifications in the experimental design allow for flexibility in the use of materials and the time required to perform the study. Furthermore, extension into additional experiments and alternative techniques are also discussed.
Bibliography:Detailed set‐up and student instruction sheets are available from the author upon request.
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ISSN:1470-8175
1539-3429
DOI:10.1002/bmb.2005.49403306426