Purification and analysis of colorful hypothetical open reading frames: An inexpensive gateway laboratory

This laboratory exercise is an inquiry‐based investigation developed around the core experiment where students, working alone or in groups, each purify and analyze their own prescreened colored proteins using immobilized metal affinity chromatography (IMAC). Here, we present reagents and protocols t...

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Bibliographic Details
Published in:Biochemistry and molecular biology education 2011-03, Vol.39 (2), p.141-144
Main Authors: DeSantis, Kara A., Reinking, Jeffrey L.
Format: Article
Language:English
Subjects:
Active Learning
affinity chromatography
Archaeal Proteins - chemistry
Archaeal Proteins - isolation & purification
Bacterial Proteins - chemistry
Bacterial Proteins - isolation & purification
Biochemistry
Chemistry
Chromatography, Affinity - methods
College Science
Color
colored proteins
Curriculum
Genetics
Humans
Inquiry
Laboratories
Laboratory Experiments
low cost
Molecular Biology
Open Reading Frames
protein purification
Science Instruction
Teaching - methods
Undergraduate
Undergraduate Students
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Summary:This laboratory exercise is an inquiry‐based investigation developed around the core experiment where students, working alone or in groups, each purify and analyze their own prescreened colored proteins using immobilized metal affinity chromatography (IMAC). Here, we present reagents and protocols that allow 12 different proteins to be purified in parallel without specialized equipment and within a 2.5‐ to 3‐hour undergraduate teaching laboratory. The visual feedback of purifying a colored biomolecule provides real‐time emphasis of the power and simplicity of recombinant DNA technology and IMAC. As presented here in its simplest form, this laboratory occupies two laboratory periods: purification followed by SDS‐PAGE analysis. As such, it can be easily inserted into the existing curriculum of a Biochemistry, Molecular Biology, Biotechnology, or even Genetics course to illustrate core concepts of central dogma and protein purification. Furthermore, the proteins in hand at the end of this 2‐week module can also be used for follow‐up experiments tailored to the needs, timeframe, and facilities available. Biochemistry and Molecular Biology Education Vol. 39, No. 2, pp. 141–144, 2011
ISSN:1470-8175
1539-3429
DOI:10.1002/bmb.20459