Administrative
Manage lab of approximately 35 employees (professors, research assistant professors, post-doctoral associates, graduate students, technical staff, and undergraduate students). Manage spending and salary distribution on 26 grants, approximately $1.5M annual budget. Place supply and equipment orders, justify all purchases, and properly charge grants. Act as liaison between office administrative (accountants and HR) and lab staff. Work with Office of Human Resources and Medical School to get hiring approval, write and position postings for new hires, interview and advise on technical staff hires. Hire, supervise, and train undergraduate student workers. Orient new employees to lab procedures, document safety training, and request UCard access to building. Schedule weekly lab meetings, keep lab calendar current. Organize lab help document (DDTLABHELP), maintain hyperlinks, keep protocols current, and recruit other responsible lab members to document their protocols.
Wetlab
Monitor lab inventory and order all supplies and equipment. Maintain service contracts on all equipment, schedule service calls, and coordinate repairs. Write and maintain research compliance protocols (IACUC, IRB, and IBC). Oversee semi-annual animal and hazardous waste inspections with various regulatory offices. Serve as lab safety and controlled substance officer. Provide annual safety and chemical training to all employees, and oversee all waste compliance in lab. Act as point-of-contact for any safety or chemical related problems and communicates with Dept of Environmental Health and Safety to resolve problems.
Scientific
Assess protein needs for muscle group. Coordinate and organize protein preps and document in mutant database. Advise lab members in molecular biology. Aid in the creation of new myosin mutants.
Research Techniques
- Molecular biology (restriction digest, ligation, plasmid purification)
- Site-directed mutagenesis
- PCR
- Cloning / subcloning
- Cell culture (E.coli, Spodoptera frugiperda, Dictyostelium discoideum)
- Sterile technique
- Protein expression (E.coli, Spodoptera frugiperda, Dictyostelium discoideum)
- Western blots
- Gel electrophoresis (SDS-PAGE, agarose, urea)
- Gel purification (DNA)
- Protein purification (native and expressed)
- Column Chromatography (anion exchange, size exclusion, affinity)
- Spectroscopic labeling (Electron Paramagnetic Resonance, Fluorescence)
- UV/Vis spectrophotometry
- Protein concentration assays (Bradford, BCA)
- ATPase activity assays (Malachite Green method)
- Fiber dissection (rabbit, scallop)
Publications
Nesmelov, Y.E., R. Agafonov, I.V. Negrashov, S. Blakely, M.A. Titus, and D.D. Thomas. 2011. Structural kinetics of myosin by transient time-resolved FRET. Proc Nat Acad Sci USA 108:1891-1896.
Agafonov R, I. V. Negrashov, Y. Tkachev, S. Blakely, M. A. Titus, D. D. Thomas, and Y. E. Nesmelov. 2009. Structural dynamics of the myosin relay helix by time-resolved EPR and FRET. Proc Nat Acad Sci USA. 106: 21625-21630.
Korman, V.L., S.E. Anderson, E. Prochniewicz, M.A. Titus, and D.D. Thomas. 2006. Structural dynamics of the actin-myosin interface by site-directed spectroscopy. J Mol Biol. 356: 1107-1117.
Nelson, W.D., S.E. Blakely, Y.E. Nesmelov, and D.D. Thomas. 2005. Site-directed spin labeling reveals a conformational switch in the phosphorylation domain of smooth muscle myosin. Proc Natl Acad Sci U S A. 102: 4000-4005. PMCID: 554790.