Marina Alekseevna Titok
Professor, Doctor of Biological Sciences.
E-mail: titokbsu.by, m_titokyahoo.com
- Vector Systems
- Extrachromosomal Genetic Elements of Bacteria
- Molecular Aspects of Evolution
- Microbial Genetics
Main directions of scientific activity:
studying the molecular-genetic organisation of extrachromosomal genetic elements of gram-positive and gram-negative bacteria and construction of vector systems for molecular cloning for the purpose of studying gene expression and creating strains-producers of biologically active compounds.
Titok M.A. was born in 1959. After graduating with honours from Belarusian State University she was working as a senior laborant and later as a junior research worker of the sector of biochemical genetic of bacteria in the research laboratory of experimental biology of Belarusian State University. Since 1991 Titok M.A. had been working as an assistant and later as an assistant professor of the Department of Genetics, since 2005 – as a professor of the Genetics Department, since 2009 – as a professor of the Department of Microbiology of BSU.
In 1991 Titok M.A. maintained a thesis and acquired the rank of Candidate of Sciences. In 1997 she get an academic status of Associate professor. Titok M.A. was granted a doctor’s degree in 2005. An academic status of Professor was awarded in 2007.
Under the direction of Professor Titok M.A. the research into studying genetic organisation of extrachromosomal genetic elements that were isolated from natural strains of gram-positive and gram-negative bacteria is carried out.
By means of comparative analysis a new theta-type plasmid of B. subtilis has been isolated and characterized. This plasmid is unique and has no analogues from the known extrachromosomal genetic elements of gram-positive and gram-negative microorganisms. It has been demonstrated that the new type of replicon is widely distributed among natural strains of B. subtilis isolated from various environmental sources on the territory of Belarus. Chromosomal determinants required for the stable maintenance of this class of extrachromosomal genetic elements have been found. A new function has been demonstrated for the genes determining the synthesis of the proteins of primosomal complex and chromosome replication initiation system. A mini-replicon of this plasmid was isolated. Based on this mini-reolicon, a vector for molecular cloning in B. subtilis was created.
The plasmids from P-9 incompatibility group of bacteria from the Pseudomonas genus were characterized. These plasmids are represented by different extrachromosomal genetic elements (R- and D-plasmids) that can be inherited and maintained in cells of different gram-negative bacteria. First of all this group of plasmids excites interest because of its ability to utilize different organic compounds. This ability permits to use bacteria that contain these plasmids as effective destructors for cleaning environment from different types of pollution. Mini-replicon of an IncP-9 group R-plasmid that has been sequenced gives reasons to study delicate mechanism of inheritance of biodegradation plasmids in the cells of homo- and heterologous hosts. The characteristic peculiarities of plasmids of the IncP-9 group which were determined: 1. Plasmids of the IncP-9 group are characterized by a wide polymorphism in molecular organization of replication initiation regions. They are the most various group of extrachromosomal genetic elements. Plasmids vary in sequences of the rep-gene and oriV. 2. The rep-gene and oriV site are sufficient for the replication of IncP-9 plasmids in P. putida, while the par-locus in cis-position is additionally required for replication in heterologous hosts E. coli because par-locus provides cell division with active distribution of plasmid copies. It means that there is a definite type of interaction between components of replicative and segregative systems that allows extrachromosomal genetic element to be maintained in foreign genetic surrounding. 3. The IncP-9 group plasmids cannot replicate in bacteria of the genus Enterobacteriaceae (Esherichia coli, Erwinia chrysanthemi, Erwinia herbicola, Salmonella typhimurium, Klebsiella aerogenes, Citobacter freundii, Serratia marcescens). The termoresistant variants of the IncP-9 group plasmid pMT2 were obtained and characterized. This feature allowed to construct a suicidal pM3 based vector and to use it for transposon mutagenesis in Enterobacteriaceae.
The main part of research was conducted and is still conducted within the bounds of research themes and programs:
1. INTAS97-1464 «Function analysis of genes on plasmids in Bacillus» (1999-2001).
2. INTAS-99-01487 «The Pseudomonas horizontal gene pool: the role of IncP-9 plasmids in the diversity of this genus» (2000-2003).
3. INTAS 01-2383 «Characterization and exploitation of plasmids controlling biodegradation of naphthalene by Pseudomonas species» (2002-2005).
4. The project of The Ministry of Education «Molecular-genetic analysis of replication systems and stable maintaining of theta-type plasmids of gram-positive and gram-negative bacteria» (2003-2005)
5. Theme 17.17 of the state program «Genetic engineering»: «To create a host-vector for cloning and expression of prokaryotic and eukaryotic genes in bacteria Bacillus subtilis» (2003-2006)
6. Student grant BSU «Creation of vector systems for gram-positive microorganisms Bacillus subtilis»
7. World Federation of Scientists, National Scholarship Program – Belarus 2004
8. Theme 13 of the state program «Biotechnology» «To isolate from natural sources strains of bacteria Bacillus subtilis that inherit different types of plasmids. To characterize main properties of plasmids and to create vectors on the basis of them for molecular cloning» (2004-2006)
9. The project of The Belarusian republican fond of fundamental research «Studying plasmids with theta-type replication that are contained into strains of Bacillus subtilis that were isolated from Belarusian soils»
10. The project of The Belarusian republican fond of fundamental research «Studying plasmids controlling biodegradation of naphthalene in strains of bacteria that were isolated from Belarusian soils» (2004-2006)
11. Theme 1.08 of the state program «New biotechnologies» «Studying genetic variety and estimating of biodegradation potential of bacteria-destructors of polycyclic aromatic hydrocarbons that were isolated from natural sources on the territory of Belarus»
12. The project of The Belarusian republican fond of fundamental research «The peculiarities of inheritance of the IncP-9 group plasmids in the cells of homo- and heterologous hosts» (2007-2009)
13 The joint Russian-Belarusian project of the Fond of fundamental research «The role of extrachromosomal and chromosomal genetic determinants in microbial degradation of polycyclic aromatic hydrocarbons» (2008-2010)
Titok M.A. Plasmids of gram-positive bacteria / ed. Fomichev Yu.K. – Minsk, BSU – 2004
Certificate of recognition:
Titok M.A., Maksimova N.P., Proculevich V.A., Fomichev Yu.K. Certificate of recognition №1599434 USSR. Plasmid DNA pMTF59 that was suggested for the transposon mutagenesis of bacteria Enterobacteriaceae // Discoveries. Inventions. -1990. -№38 – p.111
1. Titok M.A., Proculevich V.A., Maksimova N.P., Fomichev Yu.K. The use of Pseudomonas sp. M plasmid pM3 for the transposon mutagenesis of Erwinia //Mol. Gen. Microbiol. Virusol. – 1989, -№11. – p.45-48
2. Titok M.A., Lysak V.V., Kulba A.M. R-plasmids of facultative methyltrophic Pseudomonas: the characteristic of plasmid pM3 // Vestnic BSU. – s.2, №2. – 1989. – p. 42-46
3. Titok M.A., Maksimova N.P., Fomichev Yu.K. Characteristics of the R-plasmid pM3 (IncP-9) of a broad circle of hosts // Mol. Gen. Microbiol. Virusol. – 1991. - №9. – p.9-13
4. Titok M.A., Olekhnovich I.N., Evtushenkov A.N. Cloning of the REP-region of plasmids from a broad circle of pM3 hosts (IncP-9) // Mol. Gen. Microbiol. Virusol. – 1991. - №3. – p. 18-23.
5. Greated A., Titok M.A., Krasowiak R., Fairclough R.J., Thomas C.M. The replication and stable-inheritance functions of IncP-9 plasmid pM3 // Microbiol. – 2000. –Vol.146. – p.2249-2258.
6. Lotareva O.V., Nezametdinova V.Z., Fedorina E.A., Poluektova E.U., Titok M.A., Prozorov A.A. High frequency conjugative mobilization in natural strains of Bacillus subtilis, bearing a large plasmid //Genetics. – 2001. – Vol.37, №12. – p. 1598-1603
7. Lotareva O.V., Poluektova E.U., Titok M.A., Prozorov A.A. A large plasmid of a soil Bacillus subtilis strain mediates conjugative mobilization at high frequency // Reports of NAS. – 2001. – Vol.379, №1. – p.130-131.
8. Lotareva O.V., Poluektova E.U., Titok M.A., Prozorov A.A. Soil strain of Bacillus subtilis harboring a large plasmid that mediates high-frequency conjugal mobilization // Microbiol. – 2002. – Vol.71, №2. – p. 255-257.
9. Krasowiak R., Smalla K., Sokolov S., Kosheleva I.A., Sevastianovich Y.R., Titok M.A., Thomas C.M. PCR primers for detection and characterization of IncP-9 plasmids // FEMS Microbiol. Ecol. - 2002. - Vol. 42, № 2. - p. 217-225.
10. Titok M.A., Chapuis J., Selezneva Y.V., Lagodich, A.V., Prokulevich V.A., Ehriich S.D., Janniere L. Bacillus subtilis soil isolates: plasmid replicon analysis and construction of a new theta-replicating vector // Plasmid. - 2003. - Vol. 49, .N° 1. – p. 53-62.
11. Titok M.A. Application of pM3, a broad host range IncP-9 plasmid, for genetic analysis in Enterobacteriaceae // Genetics. – 2003. – Vol.47, №12. – p.1606-1611.
12. Titok M.A. Molecular genetic analysis of rep-купшщт of theta-type plasmid pBS72 of bacteria Bacillus subtilis // Reports of NAS. – 2003. – Vol.47, №4. – p.67-70
13. Titok M.A., Lagodich A.V., Selezneva Yu.V. Plasmid composition of bacteria Bacillus subtilis that were received from natural sources // Vestnic BSU. – 2003. – s.2, №3. –p.35-38
14. Lagodich A.V., Shtanyuk Ya.V., Prozorov A.A., Titok M.A. The characteristic of plasmid replication systems of natural strains of Bacillus subtilis // Mol. Biol. – 2004. –Vol.38, №3. –p.1-5
15. Titok M.A., Lagodich A.V. The characteristic of rep-region of Bacillus subtilis plasmids // News of NAS. – 2004. -№2. – p.61-65
16. Titok M.A. Genetic determinants of incompatibility of plasmid pMT2 (IncP-9) // Vestnic BSU. -2005. –s.2, №1. – p.31-36
17. Titok M.A. The temperature instability property of plasmid pMT2 (IncP-9) // Vestnic BSU. -2005. –s.2, №1. – p.26-31
18. Vasilenko S.L., Titok M.A., Maksimova N.P. Constructing of system of genetic analysis in Pseudomonas mendocina // Genetics. – 2000. –Vol.36. -№1. – p.28-34.
19. Vasilenko S.L., Titok M.A., Maksimova N.P. The optimization of mutagenesis conditions for bacteria Pseudomonas mendocina // Vestnic BSU. – 2003. - s.2, №1. – p.43-47
20. Vasilenko S.L., Maksimova N.P., Titok M.A. Conjugation mapping of Pseudomonas mendocina bacteria // Genetics. -2003. –Vol.39,№11.
21. Vasilenko S.L., Maksimova N.P., Titok M.A. Construction of a genetic map of Pseudomonas mendocina bacteria // Genetics – 2003. – Vol.39, №12
22. Levchuk A.A., Vasilenko S.L., Bulyga I.M., Titok M.A., Thomas K.M. Characteristics of natural strains of naphthalene-utilizing bacteria of the genus Pseudomonas // Izv Akad Nauk Ser Biol. -2005. -№2. – p.162-167
23. Lagodich A..V, Cherva E.A., Shtaniuk la.V., Prokulevich V.A., Fomichev lu.K., Prozorov A.A., Titok M.A. Construction of vector system for molecular cloning in Bacillus subtilis and Escherichia coli // Mol. Biol. -2005. – Vol.39, №2. – p.345-348
24. Titok M.A., Proculevich V.A., Janniere L. The influence of the host cell replication machinery on the inheritance of the pBS72 plasmid of Bacillus subtilis // Proc. NAS Belarus – 2005. – Vol.49, №3. – p.70-76
25. Titok M.A. The functional analysis of rep-region of plasmid pMT2 (IncP-9) // Proc. NAS Belarus – 2005. – Vol.49, №4. –p. 81-87
26. Sevastsyanovich Y.R., Titok M.A., Krasowiak R., Bingle L.E.H., Thomas C.M. Ability of IncP-9 plasmid pM3 to replicate in Escherichia coli is dependent on both rep and par functions. // Mol. Microbiol., -2005. - 57 (3) –p. 819-833.
27. Titok M., Suski C., Dalmais B., Ehrlich S. D., Janniere L. The replicative polymerases PolC and DnaE are required for theta replication of the Bacillus subtilis plasmid pBS72 // Microbiology (2006), 152, 1471-1478
28. Levchuk A.A., Bulyga I.M., Izmalkova T.Iu., Sevast'ianovich la.R., Kosheleva I.A., Thomas C.M., Titok M.A. Nah-plasmids of IncP-9 group from natural strains of Pseudomonas // Mol. Biol. – 2006. – Vol.40, №5. – p.835-843
29. Bulyga I.M., Titok M.A. The organization of replication initiation systems of the IncP-9 group plasmids // Reopts NAS Belarus – 2006. –Vol.5, №5 –p.63-68
30. Nezametdinova V.Z., Fedorina E.A., Poluektova E.U., Titok M.A., Prozorov A.A. Comparative characterization of the conjugation capacity and large plasmids in native strains of Bacillus subtilis from different regions of the east European plain // Microbiol. – 2007. – Vol.76,№2 – p.1-6
31. Janniere L, Canceill D, Suski C, Kanga S, Dalmais B, Lestini R, Monnier AF, Chapuis J, Bolotin A, Titok M, Le Chatelier E, Ehrlich SD. Genetic evidence for a link between glycolysis and DNA replication. // PLoSONE. – 2007 -16;2:e447.
32. Vasilenko S.L., Titok M.A. Inheritance of biodegradation plasmids in the cells of homo- and heterologous hosts // Microbiol. – 2008. –Vol.77, №1 – p.16-22
33. Sevastsyanovich Y. R., Krasowiak R., Bingle L. E. H., Haines A. S., Sokolov S. L., Kosheleva I. A., Leuchuk A. A., Titok M. A., Smalla K., Thomas C. M. Diversity of IncP-9 plasmids of Pseudomonas // Microbiology.- 2008. - V.I 54, № 10. - p. 29-41.
1. Titok M.A., Maksimova N.P. Control works for the course «Genetics» for students of correspondence studying, methodical recommendations. Minsk, BSU,1995,11p.
2. Maksimova N.P., Titok M.A. Methodical directions for the execution of yearly projects and graduate works in genetics, methodical directions. Minsk, BSU, 1995, 8 p.
3. Titok M.A., Maksimova N.P., Anochina V.S. Methodical recommendations for the course «Genetics», methodical recommendations. Minsk, BSU, 1997, 20 p.
4. Glushen S.V., Kunickaya M.P., Titok M.A. Methodical directions for practical studies for the course «Cytology and histology» methodical directions. Minsk, BSU, 1997.
5. Titok M.A., Grenev V.V. «Chromosome mapping» workbook. Minsk, BSU, 2003, 12p.
6. Titok M.A., Grenev V.V. «Linked inheritance of indications and crossingover» workbook. Minsk, BSU, 2003, 10p.
7. Titok M.A., Grenev V.V. «Interaction of genes» workbook. Minsk, BSU, 2003, 20p.
8. Glushen S.V., Grenev V.V, Kunickaya M.P., Titok M.A. «Cytology and histology» methodical directions. Minsk, BSU, 2004, 6p.
9. Titok M.A., Anochina V.S. «Evolutionary theory» methodical directions for workshops
10. Maksimova N.P., Titok M.A., Anochina V.S., Chramcova E.A., Grenev V.V., Kunickaya M.P. Workbook for genetics: manual for students of the biological faculty. Minsk, BSU, 2008, 167 p.
11. MaksimovaN.P., Titok M.A., Kunickaya M.P., Chramcova E.A., Anochina V.S. Genetics: methodical directions for laboratory works for students of the biological faculty of specialties 1-31 01 01 – Biology, 1-33 01 01 – Bioecology. – Minsk, BSU, 2008, 64p.
12. Titok M.A. Part 1 «Molecular aspects of evolution». Textbook. 183p.
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