Department of Eukaryote Gene Engineering is focused on the research directions of recombinant viral and human protein analysis, expression and assembly of proteins into virus-like particles (VLPs) as well as mechanisms of plant signaling and biosynthesis. We are using methods of gene cloning and expression in different host systems, electron microscopy, immunological and functional biochemical assays of proteins, biosynthesis and purification of essential amounts of target recombinant proteins with application area including diagnostics and therapeutics.
Research activities
- Genetic mutations of Sars-CoV2 virus detection study in Lithuania, established by the decision of the Minister of Health on 2021-02-04, no V-221 “Organizing, coordinating and investigating in-deph testing of Sars-CoV-2 in the process of detecting genetic mutations”.
- Production of recombinant viral proteins in yeast cells.
- Improvement of expression systems and yeast genetic background for efficient synthesis of viral proteins.
- Improvement of yeast protein secretion capacities.
- Identification and characterization of new polyomaviruses, parvoviruses, paramyxoviruses and hantaviruses.
- Investigation of virus-like particles (VLPs) formation in yeast cells.
- Employment of hamster polyomavirus VLPs as carriers for short and non-immunogenic protein sequences presentation on VLPs.
- Identification of new protein carriers.
- Human chaperone protein synthesis in yeast.
- Evaluation of diagnostic tests based on recombinant viral proteins.
- Synthesis of recombinant proteins in E.coli.
- Analysis and research of recombinant biopharmaceutical proteins (human cytokines, hormones, viral proteins, recombinant antibody fragments).
- Research and analysis of recombinant allergens (human protein allergens of different groups).
- Protein synthesis in plant transient expression systems.
- Research of plant anthocyanin synthesis regulation.
- Viroid studies.
- Molecular mechanisms of zebrafish Danio rerio heart regeneration.
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Main publications
2023
- Sližienė A, Plečkaitytė M, Rudokas V, Juškaitė K, Žvirblis G, Žvirblienė A. (2023). Cross-reactive monoclonal antibodies against fish parvalbumins as a tool for studying antigenic similarity of different parvalbumins and analysis of fish extracts. Molecular Immunology. doi: 10.1016/j.molimm.2023.01.001
2022
- Jiang Z, El-Brolosy MA, Serobyan V, Welker JM, Retzer N, Dooley CM, Jakutis G, Juan T, Fukuda N, Maischein HM, Balciunas D, Stainier DYR. (2022). Parental mutations influence wild-type offspring via transcriptional adaptation. Science Advances. doi: 10.1126/sciadv.abj2029.
- Kalvaityte M, Balčiūnas D. (2022). Conditional mutagenesis strategies in zebrafish. Trends in Genetics. https://doi.org/10.1016/j.tig.2022.04.007.
- Zinkevičiūtė R, Ražanskas R, Kaupinis A, Macijauskaitė N, Čiplys E, Houen G, Slibinskas R. (2022). Yeast Secretes High Amounts of Human Calreticulin without Cellular Stress. Curr Issues Mol Biol. 2022 Apr 19;44(5):1768-1787. doi: 10.3390/cimb44050122.
- Mughal FP, Bergmann AC, Huynh HUB, Jørgensen SH, Mansha I, Kesmez M, Schürch PM, Theocharides APA, Hansen PR, Friis T, Holmström MO, Ciplys E, Slibinskas R, Højrup P, Houen G, Trier NH. (2022). Characterization of Peptide Antibodies to the C-Terminal of Frameshifted Calreticulin Associated with Myeloproliferative Diseases. doi: 10.3390/ijms23126803.
- Drobysh M, Liustrovaite V, Baradoke A, Rucinskiene A, Ramanaviciene A, Ratautaite V, Viter R, Chen CF, Plikusiene I, Samukaite-Bubniene U, Slibinskas R, Ciplys E, Simanavicius M, Zvirbliene A, Kucinskaite-Kodze I, Ramanavicius A. (2022) Electrochemical Determination of Interaction between SARS-CoV-2 Spike Protein and Specific Antibodies. Int J Mol Sci. doi: 10.3390/ijms23126768.
- Kyllesbech C, Trier N, Slibinskas R, Čiplys E, Tsakiri A, Frederiksen JL, Houen G. (2022). Virus-specific antibody indices may supplement the total IgG index in diagnostics of multiple sclerosis. J Neuroimmunol. doi: 10.1016/j.jneuroim.2022.577868.
- Ratautaite V, Boguzaite R, Brazys E, Ramanaviciene A, Čiplys E, Juozapaitis M, Slibinskas R, Bechelany M, Ramanavicius A. (2022). Molecularly imprinted polypyrrole based sensor for the detection of SARS-CoV-2 spike glycoprotein. Electrochim Acta. doi: 10.1016/j.electacta.2021.139581.
- Trier NH, Valdarnini N, Fanelli I, Rovero P, Hansen PR, Schafer-Nielsen C, Čiplys E, Slibinskas R, Pociot F, Friis T, Houen G. (2022). Peptide Antibody Reactivity to Homologous Regions in Glutamate Decarboxylase Isoforms and Coxsackievirus B4 P2C. Int J Mol Sci. doi: 10.3390/ijms23084424.
- Paškevičius Š, Dapkutė V, Misiūnas A, Balzaris M, Thommes P, Sattar A, Gleba Y, Ražanskienė A. (2022). Chimeric bacteriocin S5-PmnH engineered by domain swapping efficiently controls Pseudomonas aeruginosa infection in murine keratitis and lung models. Sci Rep. doi: 10.1038/s41598-022-09865-8.
- Paškevičius Š, Gleba Y, Ražanskienė A. (2022). Stenocins: Novel modular bacteriocins from opportunistic pathogen Stenotrophomonas maltophilia. J Biotechnol. doi: 10.1016/j.jbiotec.2022.04.006.
- Lučiūnaitė A, Dalgėdienė I, Žilionis R, Mašalaitė K, Norkienė M, Šinkūnas A, Gedvilaitė A, Kučinskaitė-Kodzė I, Žvirblienė A. (2022). Activation of NLRP3 Inflammasome by Virus-Like Particles of Human Polyomaviruses in Macrophages. Frontiers in Immunology. doi: 10.3389/fimmu.2022.831815.
2021
- Jandrig B, Krause H, Zimmermann W, Vasiliunaite E, Gedvilaite A, Ulrich RG. (2021). Hamster Polyomavirus Research: Past, Present, and Future. Viruses 13(5):907. doi: 10.3390/v13050907.
- Eiden M, Gedvilaite A, Leidel F, Ulrich RG, Groschup MH. (2021). Vaccination with Prion Peptide-Displaying Polyomavirus-Like Particles Prolongs Incubation Time in Scrapie-Infected Mice. Viruses 13(5):811. doi: 10.3390/v13050811.
- Houen G, Højrup P, Ciplys E, Gaboriaud C, Slibinskas R. (2021). Structural Analysis of Calreticulin, an Endoplasmic Reticulum-Resident Molecular Chaperone. Prog Mol Subcell Biol. 2021;59:13-25. doi: 10.1007/978-3-030-67696-4_2.
- Plikusiene I, Maciulis V, Ramanaviciene A, Balevicius Z, Buzavaite-Verteliene E, Ciplys E, Slibinskas R, Simanavicius M, Zvirbliene A, Ramanavicius A.J. (2021). Evaluation of kinetics and thermodynamics of interaction between immobilized SARS-CoV-2 nucleoprotein and specific antibodies by total internal reflection ellipsometry. Colloid Interface Sci. 2021 Jul 15;594:195-203. doi: 10.1016/j.jcis.2021.02.100. Epub 2021 Mar 10.
- Bulavaitė A, Maier T, Pleckaityte M. (2021) Discrimination of Gardnerella Species by Combining MALDI-TOF Protein Profile, Chaperonin cpn60 Sequences, and Phenotypic Characteristics. Pathogens. 2021 Mar 1;10(3):277. doi: 10.3390/pathogens10030277. PMID: 33804525; PMCID: PMC7998583.
- Čiplys E, Paškevičius T, Žitkus E, Bielskis J, Ražanskas R, Šneideris T, Smirnovas V, Kaupinis A, Tester DJ, Ackerman MJ, Højrup P, Michalak M, Houen G, Slibinskas R. (2021). Mapping human calreticulin regions important for structural stability. Biochim Biophys Acta Proteins Proteom. 1869(11):140710. doi: 10.1016/j.bbapap.2021.140710.
- McNabb L, Andiani A, Bulavaite A, Zvirbliene A, Sasnauskas K, Lunt R. (2021). Development and validation of an IgM antibody capture ELISA for early detection of Hendra virus. J Virol Methods. 21;298:114296. doi:10.1016/j.jviromet.2021.114296.
- Dudas G, Hong SL, Potter BI, Calvignac-Spencer S, Niatou-Singa FS, Tombolomako TB, Fuh-Neba T, Vickos U, Ulrich M, Leendertz FH, Khan K, Huber C, Watts A, Olendraitė I, Snijder J, Wijnant KN, Bonvin AMJJ, Martres P, Behillil S, Ayouba A, Maidadi MF, Djomsi DM, Godwe C, Butel C, Šimaitis A, Gabrielaitė M, Katėnaitė M, Norvilas R, Raugaitė L, Koyaweda GW, Kandou JK, Jonikas R, Nasvytienė I, Žemeckienė Ž, Gečys D, Tamušauskaitė K, Norkienė M, Vasiliūnaitė E, Žiogienė D, Timinskas A, Šukys M, Šarauskas M, Alzbutas G, Aziza AA, Lusamaki EK, Cigolo JM, Mawete FM, Lofiko EL, Kingebeni PM, Tamfum JM, Belizaire MRD, Essomba RG, Assoumou MCO, Mboringong AB, Dieng AB, Juozapaitė D, Hosch S, Obama J, Ayekaba MO, Naumovas D, Pautienius A, Rafaï CD, Vitkauskienė A, Ugenskienė R, Gedvilaitė A, Čereškevičius D, Lesauskaitė V, Žemaitis L, Griškevičius L, Baele G. (2021). Emergence and spread of SARS-CoV-2 lineage B.1.620 with variant of concern-like mutations and deletions. Nat Commun. 2021 Oct 1;12(1):5769. doi: 10.1038/s41467-021-26055-8.
- Bergmann AC, Kyllesbech C, Slibinskas R, Ciplys E, Højrup P, Trier NH, Houen G. (2021). Epitope Mapping of Monoclonal Antibodies to Calreticulin Reveals That Charged Amino Acids Are Essential for Antibody Binding. Antibodies (Basel). 2021 Aug 4;10(3):31. doi: 10.3390/antib10030031.
- Lazutka J, Simutis K, Matulis P, Petraitytė-Burneikienė R, Kučinskaitė-Kodzė I, Simanavičius M, Tamošiunas PL. (2021) Antigenicity study of the yeast-generated human parvovirus 4 (PARV4) virus-like particles. Virus Res. 2021 Jan 15;292:198236. doi: 10.1016/j.virusres.2020.198236.
- Kaliniene L, Noreika A, Kaupinis A, Valius M, Jurgelaitis E, Lazutka J, Meškienė R, Meškys R. (2021). Analysis of a Novel Bacteriophage vB_AchrS_AchV4 Highlights the Diversity of Achromobacter Viruses. Viruses. 2021 Feb 27;13(3):374. doi: 10.3390/v13030374.
- Kubickova B, Schenk JA, Ramm F, Markuškienė K, Reetz J, Dremsek P, Tamosiunas PL, Cepulyte L, Trinh HA, Scholz J, Memczak H, Hovestädt M, Ryll R, Petraityte-Burneikiene R, Corman VM, Andersson A, Becher D, Groschup MH, Kubick S, Sellrie F, Johne R, Ulrich RG. (2021). A broadly cross-reactive monoclonal antibody against hepatitis E virus capsid antigen. Appl Microbiol Biotechnol. 2021 Jun;105(12):4957-4973. doi: 10.1007/s00253-021-11342-7.
- Vilys L, Peciuliene I, Jakubauskiene E, Zinkeviciute R, Makino Y, Kanopka A. (2021). U2AF - Hypoxia-induced fas alternative splicing regulator. Exp Cell Res. 2021 Feb 1;399(1):112444. doi: 10.1016/j.yexcr.2020.112444.
2020
- Starkevič, P., Ražanskienė, A., Starkevič, U., Kazanavičiūtė, V., Denkovskienė, E., Bendokas, V., Šikšnianas, T., Rugienius, R., Stanys, V., and Ražanskas, R. (2020). Isolation and Analysis of Anthocyanin Pathway Genes from Ribes Genus Reveals MYB Gene with Potent Anthocyanin-Inducing Capabilities. Plants 9, 1078. 10.3390/plants9091078
- Noreika, A., Meškys, R., Lazutka, J., and Kaliniene, L. (2020). Complete genome sequence of Buttiauxella phage vB_ButM_GuL6. Arch Virol 165, 2685–2687. 10.1007/s00705-020-04780-7
- Lazutka, J., Simutis, K., Matulis, P., Petraitytė-Burneikienė, R., Kučinskaitė-Kodzė, I., Simanavičius, M., and Tamošiunas, P.L. (2020). Antigenicity study of the yeast-generated human parvovirus 4 (PARV4) virus-like particles. Virus Research 198236. 10.1016/j.virusres.2020.198236
- Špakova, A., Dalgėdienė, I., Insodaitė, R., Sasnauskienė, A., Žvirblienė, A., and Petraitytė-Burneikienė, R. (2020). vB_EcoS_NBD2 bacteriophage-originated polytubes as a carrier for the presentation of foreign sequences. Virus Research 290, 198194. 10.1016/j.virusres.2020.198194
- Plikusiene, I., Balevicius, Z., Ramanaviciene, A., Talbot, J., Mickiene, G., Balevicius, S., Stirke, A., Tereshchenko, A., Tamosaitis, L., Zvirblis, G., et al. (2020). Evaluation of affinity sensor response kinetics towards dimeric ligands linked with spacers of different rigidity: Immobilized recombinant granulocyte colony-stimulating factor based synthetic receptor binding with genetically engineered dimeric analyte derivatives. Biosensors and Bioelectronics 156, 112112. 1016/j.bios.2020.112112
- Denkovskienė, E*., Paškevičius, Š*., Stankevičiūtė, J., Gleba, Y., and Ražanskienė, A. (2020). Control of T-DNA Transfer from Agrobacterium tumefaciens to Plants Based on an Inducible Bacterial Toxin-Antitoxin System. MPMI MPMI-03-20-0067. 10.1094/MPMI-03-20-0067-R
- Vilmane, A., Terentjeva, A., Tamosiunas, P.L., Suna, N., Suna, I., Petraityte-Burneikiene, R., Murovska, M., Rasa-Dzelzkaleja, S., and Nora-Krukle, Z. (2020). Human Parvoviruses May Affect the Development and Clinical Course of Meningitis and Meningoencephalitis. Brain Sciences 10, 339. 10.3390/brainsci10060339
- Houen, G., Heiden, J., Trier, N.H., Draborg, A.H., Benros, M.E., Zinkevičiūtė, R., Petraitytė-Burneikienė, R., Ciplys, E., Slibinskas, R., and Frederiksen, J.L. (2020). Antibodies to Epstein-Barr virus and neurotropic viruses in multiple sclerosis and optic neuritis. Journal of Neuroimmunology 346, 577314. 10.1016/j.jneuroim.2020.577314
- Drewes, S., Jeske, K., Straková, P., Balčiauskas, L., Ryll, R., Balčiauskienė, L., Kohlhause, D., Schnidrig, G.-A., Hiltbrunner, M., Špakova, A., Insodaitė R, Petraitytė-Burneikienė R, et al. (2020). Identification of a novel hantavirus strain in the root vole (Microtus oeconomus) in Lithuania, Eastern Europe. Infection, Genetics and Evolution 104520. 10.1016/j.meegid.2020.104520
- Bulavaitė, A., Dalgediene, I., Michailoviene, V., and Pleckaityte, M. (2020). Type II Restriction-Modification System from Gardnerella vaginalis ATCC 14018. Pathogens 9, 703. 10.3390/pathogens9090703
- Mickiene, G*., Dalgėdienė, I., Zvirblis, G., Dapkunas, Z., Plikusiene, I., Buzavaite-Verteliene, E., Balevičius, Z., Rukšėnaitė, A., and Pleckaityte, M. (2020). Human granulocyte-colony stimulating factor (G-CSF)/stem cell factor (SCF) fusion proteins: design, characterization and activity. PeerJ 8, e9788.
Projects
Research projects:
- Research Council of Lithuania. Investigation of lactis mutations conferring enhanced secretion phenotype and generation of yeast strains for supersecretion of recombinant proteins (No. S-MIP-17-88). A. Gedvilaitė. 2017–2020.
- LSC project for activity according action 2015-LMT-K-718 “Scientific research for advanced investigators“ (Smart specialization), No. 01.2.2-LMT-K-718-01-0008 ,,New technologies for construction of recombinant allergens“, G. Žvirblis, 2018-2021.
- Research Council of Lithuania/ European Social Fund. Genetic and molecular studies of the role of Tbx5a role in heart regeneration) (No. Nr. 09.3.3-LMT-K-712-17-0014) D. Balčiūnas 2020-2023.
- Research Council of Lithuania/ European Social Fund. Screening for new methods for treatment of neurodegenerative diseases (No. 2.2-LMT-K-718-03-0021). Dr. E. Čiplys. 2020-2023.
- “InnoVouchers” (Agency for Science, Innovation and Technology), contract with UAB Imunodiagnostika “Development of recombinant allergen producents”. 2021. Rasa Petraitytė-Burneikienė.
Contracts:
- Abcam Ltd, United Kingdom,
- Arc Dia International Oy Ltd, Finland,
- UAB Profarma, Lithuania,
- UAB Baltymas, Lithuania.
- UAB Imunodiagnostika, Lithuania.
Collaborations:
- Friedrich-Loeffler-Institut Bundesforschungsinstitut für Tiergesundheit, Federal Research Institute for Animal Health OIE Collaborating Centre for Zoonoses in Europe (Germany)
- Department of Virology, University of Freiburg (Germany)
- Max Planck Institute for Molecular Plant Physiology (Germany)
- Institute of Virology, Slovak Academy of Sciences (Slovakia)
- Kirchenstein Institute of Microbiology and Virology, Riga Stradins University (Latvia)
- Department of Medical Research, Mackay Memorial Hospital, (Taiwan)
Achievements
- Identified six new common and bank voles', yellow-necked mouse's, rat's and shrews’ polyomaviruses.
- Large collection of more than 40 different virus like particles (VLPs) derived from various polyomavirus (all 13 human polyomavirus, rodents, avian and other host polyomavirus) VP1 proteins and papillomavirus L1 proteins (HPV6, 16, 18, 31, 33), porcine circovirus type 2 capsid proteins.
- Proteins of measles, mumps, rubella, parainfluenza viruses (1-4), hantaviruses, porcine parvovirus, human bocaviruses (1-4), human parvovirus 4, human metapneumovirus, hepatitis E and bovine Schamallenberg virus were produced in yeast cells.
- Serological diagnostic tests were developed for detection of viral-specific antibodies in human serum and saliva samples.
- Commercially available Microiimune (UK) measles and mumps diagnostic tests are based on the proteins developed in the department.
- Human chaperons (calreticulin and BiP) were produced in yeast cells.
- Yeast expression systems for construction and production of hamster polyomavirus VP1 and VP1/VP2 VLPs as carriers harbouring non immunogenic peptides or complex molecules of interest.
- Genetic engineered yeast strain collection for recombinant protein production.
- 2003 – Lithuanian Science Prize in the Biomedicine Sciences area.
- Patent: Zvirblienė A., Gedvilaitė A., Ulrich, R., Sasnauskas. Process for the production of monoclonal antibodies using chimeric VLPs. US Patent No.:US7,919,314 B2. Apr. 5, 2011.
- Patent application: Čiplys E., Slibinskas R., Sasnauskas K., Michalak M., Gold L-I. Generation of native recombinant secreted human endoplasmic reticulum chaperones by using their native signal sequences in yeast expression systems. WO 2014/011723 A1.
Emploees
| Employee | Position | Contacts | Publications | |
| Dr. Rasa Petraitytė-Burneikienė | Head of the Departament, Senior Researcher |
Ph.: +370 5 2234421, Email: Saulėtekio al. 7, C461 |
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| Edita Bakūnaitė | Research Asisstant | Ph.: +370 5 2239429 Email: Saulėtekio al. 7, C449 |
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| Dr. Darius Balčiūnas | Research Professor | Email: Saulėtekio al. 7 |
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| Dr. Aistė Bulavaitė | Researcher | Ph.: +370 5 2239431 Email: Saulėtekio al. 7, C453 |
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| Dr. Evaldas Čiplys | Senior Researcher | Ph.: +370 5 2239432 Email: Saulėtekio al. 7, C455 |
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| Dr. Alma Gedvilaitė | Research Professor | Ph.: +370 5 2234359 Email: Saulėtekio al. 7, C467 |
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| Dr. Justas Lazutka | Researcher | Ph.: +370 5 2239435 Email: Saulėtekio al. 7, C449 |
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| Dr. Milda Norkienė | Researcher | Ph.: +370 5 2239434 Email: Saulėtekio al. 7, C463 |
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| Juta Rainytė | Research Asisstant | PH.: Email: Saulėtekio al. 7 |
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| Dr. Raimundas Ražanskas | Researcher | Ph.: +370 5 2239439 Email: Saulėtekio al. 7, V419 |
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| Eglutė Rudokienė | Senior Chemist | Ph.: +370 5 2234370 Email: Saulėtekio al. 7, V417 |
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| Dr. Rimantas Slibinskas | Research Professor | Ph.: +370 5 2239432 Email: Saulėtekio al. 7, C455 |
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| Dr. Paulius Lukas Tamošiūnas | Researcher | Ph.: +370 5 2234359 Email: Saulėtekio al. 7, V421 |
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| Dr. Mindaugas Zaveckas | Researcher | Ph.: +370 5 2239431 Email: Saulėtekio al. 7, C453 |
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| Dr. Danguolė Žiogienė | Researcher | Ph.: +370 5 2239434 Email: Saulėtekio al. 7, C463 |
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| Eimantas Žitkus | Research Asisstant | Ph.: +370 5 2239432 Email: Saulėtekio al. 7, C455 |
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| Postdoctoral Researcher: | ||||
| Dr. Jaroslav Denkovskij | Email: Saulėtekio al. 7, C453 |
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| PhD students: | ||||
| Laima Čepulytė | Email: | |
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| Miglė Kalvaitytė | Email: | |
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| Laimis Silimavičius | Email: | |
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| Emilija Vasiliūnaitė | Email: | |
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| Arūnė Verbickaitė | Email: | |
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| Technicians: | ||||
| Danutė Jaskytė | Email: | |||
| Olga Jančevskaja | Email: | |||
| Evaldas Lekavičius | Email: | |||
| Emeritus: | ||||
| Prof. habil. Dr. Kęstutis Sasnauskas | Email: | |
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