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Address
Zámek 136, CZ 373 33 Nové Hrady
Phone: +420 389 033 815
ecsefalvay@nh.cas.cz
Ing. Eva Cséfalvay, PhD.
Position
Head of the molecular biology unit
Zámek 136, CZ 373 33 Nové Hrady
Phone: +420 389 033 815
ecsefalvay@nh.cas.cz
Current projects
Eva Csefalvay is interested in the fundamental basis of the cleaving mechanism of Type I restriction-modification enzymes. Recently, she has characterized the DNA ends produced by EcoKI, EcoAI and EcoR124I, members of the Type IA, IB and IC families, respectively, by cloning and sequencing of restriction products from reactions with a plasmid DNA substrate containing a single recognition site for each enzyme (REF). The Type I restriction-modification enzymes recognize specific DNA sequences, but cleave DNA at non-specific sites far from the recognition site in a reaction that is associated with ATP-dependent DNA translocation. Although the mode of action of these enzymes has been extensively studied, it still remains elusive. Her research perspective is to gain deeper insight into the mechanism of both the DNA translocation and the DNA cleavage reactions of Type I enzymes. Recently, she suceeded in the determination of the HsdR motor subunit of the type I restriction modification system EcoR124I in collaboration with the Kuta Smatanova lab. Metods used by Csefalvay: DNA (genomic, plasmid,) and RNA isolation (soil bacteria, E. coli), DNA isolation from soil, Plasmid visualization (alkaline lyses), Cloning of plasmid DNA, PCR, sequencing PCR, RT- PCR, Real Time PCR-quantification of mRNA, Digoxigenin-labeling of DNA, southern blotting and DNA hybridization, Analysis of nucleotide and protein sequence, Expression of proteins in E. coli, Enzyme assays (catechol dioxygenases, gentisate dioxygenase), Resting cells assay, Analytical methods- FPLC, HPLC (chlorobenzoates, chlorocatechols, etc.), GS (PCBs), Biparental mating (plate, filter mating), DGGE analysis.
Institute publications in impacted journals
- Vitali Bialevich, Dhiraj Sinha, Katsiaryna Shamayeva, Alena Guzanova, David Řeha, Eva Csefalvay, Jannette Carey, Marie Weiserova, Rüdiger H Ettrich (2017) The helical domain of the EcoR124I motor subunit participates in ATPase activity and dsDNA translocation.. In: PeerJ. Vol 5, pp. 01-.
- Sinha D., Shamayeva K., Ramasubramani V., Reha D., Bialevich V., Khabiri M., Guzanova A., Milbar N., Weiserova M., Csefalvay E., Carey J., Ettrich R. (2014) Interdomain communication in the endonuclease/motor subunit of type I restriction-modification enzyme EcoR124I.. In: Journal of Molecular Modeling. Vol 20 (7), pp. 2334-.
- Csefalvay E, Lapkouski M, Komarek O (2009) Expression, purification and preliminary crystallization study of RpaC protein from Synechocystis sp PCC6803. In: PHOTOSYNTHETICA. Vol 47 (3), pp. 355-362.
- Lapkouski M., Panjikar S., Janscak P., Kuta Smatanova I., Carey J., Ettrich R., Csefalvay E. (2009) Structure of the motor subunit of type I restriction-modification complex EcoR124I. In: Nat Struct Mol Biol. Vol 16, pp. 94-95.
- Lapkouski M, Panjikar S, Smatanova IK, Csefalvay E (2007) Purification, crystallization and preliminary X-ray analysis of the HsdR subunit of the EcoR124I endonuclease from Escherichia coli. In: ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS. Vol 63 (Part 7), pp. 582-585.