Difference between revisions of "Jasmine Davila Chem430 F16"
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==Proposed Research Project== | ==Proposed Research Project== | ||
| − | === | + | ===IscR Purification from ''Escherichia coli.'' for AFM Study=== |
===General Information=== | ===General Information=== | ||
:Advisor: Laura Moore | :Advisor: Laura Moore | ||
| − | :Other research student collaborators: | + | :Other research student collaborators: none |
| − | :Other Research Collaborators: | + | :Other Research Collaborators: none |
===Proposal=== | ===Proposal=== | ||
| + | Iron sulfur cluster regulator (IscR) is an Fe-S protein that functions as a transcriptional regulator of Fe-S production and other Fe-S protein encoding genes in Escherichia coli (E. coli.). IscR controls the expression of more than 40 genes in E. coli (1). The purification of IscR for studying its many properties in E. coli has been done many times before. | ||
| + | Recently, Atomic Force Microscopy Microscope (AFM) has provided a means of studying receptor ligand interactions that were once not available to us. The NTA-His6 system has shown promise of being an efficient and powerful platform for the stable oriented immobilization of proteins in AFM single-molecule studies (2). We plan to use His-tag sequencing to bind our IscR protein and DNA to a nickel platform and investigate the binding strength of IscR to DNA using AFM techniques. | ||
===Instruments to be used=== | ===Instruments to be used=== | ||
| + | AFM (atomic force microscope) | ||
| + | Sonicator | ||
| + | Ultra Centrifuge | ||
| + | Nickel Column | ||
| + | SDS PAGE | ||
| + | |||
===References (2 minimum)=== | ===References (2 minimum)=== | ||
| + | 1 Schwartz CJ, Giel JL, Patschkowski T, Luther C, Ruzicka FJ, Beinert H, Kiley PJ. IscR, an Fe-S cluster-containing transcription factor, represses expression of Escherichia coli genes encoding Fe-S cluster assembly proteins. Proc Natl Acad Sci USA 2001; 98:14895-900 [PubMed: 11742080] | ||
| + | 2 Verbelen, C., Gruber, H. J., & Dufrêne, Y. F. (2007). The NTA-His6 bond is strong enough for AFM single-molecular recognition studies.Journal of Molecular Recognition: JMR, 20(6), 490–494. doi:10.1002/jmr.833 | ||
===Research pledge=== | ===Research pledge=== | ||
I, Jasmine Davila, have read the Chem/Bioc 430 course syllabus and understand the general structure and expectations of the research program. The above material was prepared after consultation, and in conjunction with my research advisor. | I, Jasmine Davila, have read the Chem/Bioc 430 course syllabus and understand the general structure and expectations of the research program. The above material was prepared after consultation, and in conjunction with my research advisor. | ||
Revision as of 02:36, 29 September 2016
SChemistry/Biochemistry Research 430
- Fall 2016
- Jasmine Davila
- Senior Biochemistry Major
Research Times
Tuesday 8-12
- section 01
Proposed Research Project
IscR Purification from Escherichia coli. for AFM Study
General Information
- Advisor: Laura Moore
- Other research student collaborators: none
- Other Research Collaborators: none
Proposal
Iron sulfur cluster regulator (IscR) is an Fe-S protein that functions as a transcriptional regulator of Fe-S production and other Fe-S protein encoding genes in Escherichia coli (E. coli.). IscR controls the expression of more than 40 genes in E. coli (1). The purification of IscR for studying its many properties in E. coli has been done many times before. Recently, Atomic Force Microscopy Microscope (AFM) has provided a means of studying receptor ligand interactions that were once not available to us. The NTA-His6 system has shown promise of being an efficient and powerful platform for the stable oriented immobilization of proteins in AFM single-molecule studies (2). We plan to use His-tag sequencing to bind our IscR protein and DNA to a nickel platform and investigate the binding strength of IscR to DNA using AFM techniques.
Instruments to be used
AFM (atomic force microscope) Sonicator Ultra Centrifuge Nickel Column SDS PAGE
References (2 minimum)
1 Schwartz CJ, Giel JL, Patschkowski T, Luther C, Ruzicka FJ, Beinert H, Kiley PJ. IscR, an Fe-S cluster-containing transcription factor, represses expression of Escherichia coli genes encoding Fe-S cluster assembly proteins. Proc Natl Acad Sci USA 2001; 98:14895-900 [PubMed: 11742080] 2 Verbelen, C., Gruber, H. J., & Dufrêne, Y. F. (2007). The NTA-His6 bond is strong enough for AFM single-molecular recognition studies.Journal of Molecular Recognition: JMR, 20(6), 490–494. doi:10.1002/jmr.833
Research pledge
I, Jasmine Davila, have read the Chem/Bioc 430 course syllabus and understand the general structure and expectations of the research program. The above material was prepared after consultation, and in conjunction with my research advisor.