Wlodek Mandecki, Ph.D.
Ribosome as Molecular Machine for Acquiring Sequence Information
The lab works on a method for acquiring sequence data from single nucleic acid molecules. The approach involves a fluorescence resonance energy transfer assay (FRET) based on molecules involved in protein biosynthesis. The fluorescence signal is acquired from single molecules using a fluorescence correlation spectroscope in several configurations, including measurements in solution and on surfaces. The project's goals are to: (i) perform site-directed labeling with a fluorescent dye and quencher; (ii) optimize the FRET assay; (iii) construct a synthetic template and demonstrate the performance of the system on this template; (iv) investigate nanostructures capable of enhancing fluorescence; (v) study the behavior of single molecules in the system; and (vi) demonstrate the capability of the system to acquire high volumes of sequence data. The method once fully developed will allow fast analyses of many types of nucleic acids. The project is funded by the NIH program on the "Revolutionary Genome Sequencing Technologies - the $1000 Genome".
EF-Tu (green) interacts with tRNA (pink) on the ribosome (not shown). Generated in PyMOL from data in 1mj1.pdb file. In addition, Dr. Mandecki's research interests include the mechanisms of frameshifting in ribosomal translation, phage display, protein structure and function, and innovative techniques in nucleic acid and protein analysis.
The project is a collaboration between three investigators at the Department of Microbiology and Molecular Genetics of New Jersey Medical School:
as well as the following institutions and investigators:
University of Pennsylvania:
University of North Texas Health Sciences Center
- Lin X, Flint J, Azaro M, Coradetti T, Kopacka W, Streck D, Wang Z, Dermody J, and Mandecki W (2007) Microtransponder-based multiplex assay for genotyping cystic fibrosis. Clin Chem 53: 1372-1376
- Mandecki W, Ardelt B, Coradetti T, Davidowitz H, Flint J, Huang Z, Kopacka W, Lin X, Wang Z, and Darzynkiewicz Z (2006) Microtransponders, the miniature RFID electronic chips, as platforms for cell growth in cytotoxicity assays. Cytometry Part A 69A:1097-1105
- Song, L., Mandecki, W., Goldman, E. (2003) Expression in E.coli of non-ORF sequences isolated from phage display: upstream start and stop required for translation reinitiation. FASEB J. 17: 1674-1681.
- Zemsky, J., Mandecki W., and Goldman, E. (2002) Genetic analysis of translation in the -1 frame of an unusual non-ORF sequence isolated from phage display. Gene Expression 10: 109-114.
- Goldman, E., Korus, M. and Mandecki, W. (2000) Efficiencies of translation in three reading frames of unusual non ORF sequences isolated from phage display. FASEB J. 14: 603-611.