Jasmine Davila Chem430 F16

SChemistry/Biochemistry Research 430

Fall 2016

Jasmine Davila

Senior Biochemistry Major

Research Times

Tuesday 8-12

Proposed Research Project

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 (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.