|
|
| Home > JCE Print > Journal of Chemical Education > Issues >
1999
>
March
> |
|
In the Laboratory
|
|
Concepts in Biochemistry
|
|
Recombinant Green Fluorescent Protein Isoforms: Exercises To Integrate Molecular Biology, Biochemistry, and Biophysical Chemistry
|
Barry W. Hicks
United States Air Force Academy, Department of Chemistry, USAF Academy, CO 80840
|
|
March 1999
Vol. 76 No. 3
p. 409
|
|
|
|
|
|
|
| Abstract |
|
The laboratory exercises described here can integrate fundamental concepts in biochemistry with the theoretical and applied principles of fluorescence spectroscopy. Using common techniques of molecular biology, students transfect Escherichia coli with cloned DNA (cDNA) plasmids encoding for various isoforms of the green fluorescent protein (GFP) from the jellyfish Aequorea victoria. The different GFP isoforms can then be purified and characterized. GFP functions as a fluorophore. The gene encoding the protein has been cloned, and cDNAs encoding different mutant isoforms of the protein are commercially available. The GFP cDNA can be used to give students insight into the relationship between protein structure and function by examining the effects of amino acid substitutions on the spectral properties of the fluorophore. Moreover, a wide range of chemical topics can be integrated into these experiments with different emphasis, depending upon the instructor. Possible areas of emphasis include DNA replication, RNA synthesis, protein synthesis, protein structure, recombinant DNA technology (biochemistry), the chemistry involved in production of the fluorophore and its structural properties (organic chemistry), the principles of fluorescence spectroscopy (physical chemistry), and the design and operation of a spectrofluorimeter (analytical chemistry). While ideally suited for senior-level undergraduate biochemistry exercises, these experiments could be adapted for high school or for lower-level biochemistry classes. How the experiments serve as the foundation for project-oriented laboratory exercises is also discussed.
|
|
| More Information |
 Citation |
Hicks, Barry W. J. Chem. Educ. 1999 76 409.
|
Keywords |
Proteins / Peptides; Luminescence; Molecular Properties / Structure; Biochemistry
|
History |
Created:
Last Updated: |
June 15, 1999
June 22, 2005
|
|
|
|
|
|
| Home > JCE Print > Journal of Chemical Education > Issues >
1999
>
March
> Page
409
|
|
|
| JCE HS CLIC |
|
Our Secondary School editors work hard to distill all the JCE materials to produce a fraction of particular interest to high school teachers. We call it CLIC.
|
| Contributions Welcome |
| JCE welcomes your submission |
| Advertisers |
| In recent years we have worked hard to better match our advertisers with our readers. When shopping for chemistry education materials, visit our advertisers' WWW sites first. |
| Be An Ambassador |
| Take JCE along on your outreach missions. Copies of the Journal, guest access to JCE Online, our publications catalog, and more are available for your participants. |
|
