Jasmine Davila Chem430 F16
SChemistry/Biochemistry Research 430
- Fall 2016
- Jasmine Davila
- Senior Biochemistry Major
Research Times
Tuesday 8-12
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 is a well known technique. Recently, Atomic Force Microscopy (AFM) has provided a means of studying receptor ligand interactions that were once not available to us. Using the NTA-His6 system as a platform for the isolated protein and DNA has shown promise of being an efficient way to study desired single-molecule interactions in a stable manner with AFM (2). We thus 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.