Lab of Genetic Engineering

Meet our Team

Meet our Team

lazarev

Head of laboratory

Vassili N.Lazarev

DSc, Associate Professor

Phone
+7 499 246-4665

0202

Valentin A. Manuvera, Senior Researcher, PhD

Dmitry A. Shirokov, Senior Researcher

Nadezhda F. Polina, PhD, Researcher

Daria D. Kharlapmieva, PhD, Researcher

Pavel A. Bobrovsky, Junior Researcher

Alexei S. Kurdyumov, Lab Assistant

Elena V. Alexeeva, Lab Assistant

Alexandra M. Belova, Lab Assistant

Lyudmila S. Boldyreva, Lab Assistant

Ekaterina N. Grafskaya, PhD student

Vassili K. Dimitriev, Student

Daniil A. Kornilov, Student

Olga A. Miroshina, Student

About our work

Scientific interests and research areas

  • System biology of the medicinal leech. The search for new antimicrobial peptides and thrombolytics.
  • The development of genetic therapy for mycoplasmosis and chlamydiosis using genes of antimicrobial peptides.
  • The study of mechanisms of chlamydial intracellular parasitism.
  • Cloning and expression of genes of recombinant proteins in heterologous systems. Development of methods for the isolation and purification of recombinant proteins

Theses researches

2003. Shavkunov Alexander Sergeyevich. Thesis for a scientific degree of a candidate of biological sciences by specialty 03.01.04 — Biochemistry "A study of sterol-14a-demethylase Mycobacterium tuberculosis genetic polymorphism and its expression in Escherichia coli"

2007. Kostryukova Elena Sergeyevna. Thesis for a scientific degree of a candidate of biological sciences by specialty 03.01.04 — Biochemistry, 03.01.03 — Molecular Biology "Cloning and expression of genes encoding Chlamydia trachomatis’s inclusion membrane proteins IncA-IncG"

2008. Basovsky Yuri Ivanovich. Thesis for a scientific degree of a candidate of biological sciences by specialty 03.01.04 — Biochemistry, 03.01.03 — Molecular Biology "Structural and functional characterization of Chlamydia trachomatis’s inclusion membrane proteins"

2010. Lazarev Vassili Nickolaevich. Thesis for a scientific degree of a doctor of biological sciences by specialty 03.01.03 — Molecular Biology "The new strategy of using genes of antimicrobial peptides from the arthropods venom as gene therapy agents"

2016. Polina Nadezhda Fedorovna. Thesis for a scientific degree of a candidate of biological sciences by specialty 03.01.04 — Biochemistry "Biological activity of antimicrobial peptides from the Lachesana tarabaevi spider‘s venom for Chlamydia trachomatis infection model"

2016. Kharlampieva Daria Dmitrievna. Thesis for a scientific degree of a candidate of biological sciences by specialty 03.01.04 — Biochemistry "Recombinant fragilyzins from Bacteroides fragilis, and the study of their biological activity"

 

Research grants

RFBR grant # 16-34-00480 "Investigation of the isopeptidase activity mechanism of the destabilase from the medicinal leech." in the 2016-2017 biennium.

RFBR grant # 16-34-00936 "Searching for novel antimicrobial peptides that are active against intracellular bacteria, and investigation of their action mechanism" in the 2016-2017 biennium.

RSF Grant # 14-14-00696 "System analysis of salivary glands secretion of medicinal leeches of various species (Hirudo medicinalis, H. verbana, H.orientalis) for the directed search of new antimicrobial proteins and peptides" for 2014-2016 triennium.

Scientific contacts

  • Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, laboratory of neuroreceptors and neuroregulators
    http://www.ibch.ru/structure/groups/nrnr
    The direction of joint research: preparation of cDNA libraries of genes from the venom of spiders, high-throughput sequencing, search and study of new activity of membrane-active peptides.
  • Institute for Veterinary Medical Research, Centre for Agricultural Research, Hungarian Academy of Sciences
    http://www.vmri.hu/atki_a.htm
    The direction of joint research: the development of gene-therapeutic treatment of mycoplasmosis in poultry.
  • Moscow State University, Department of Human and Animal Physiology
    http://human.bio.msu.ru
    The direction of joint research: the study of biologically active substances produced by the medicinal leech.
  • The National Center for Biotechnology of the Republic of Kazakhstan, laboratory of immunochemistry and immunobiotechnology
    http://www.ru.biocenter.kz/ncb-labs/lii.html
    The direction of joint research: development of enzyme immunoassay test kits for the Treponema pallidum diagnosis in human and FMD virus in farm animals.

Patents

  • RF Patent for invention № 2520083 "The method for determining of activity of transcription factors"
  • RF Patent for invention № 2556823 "The method of evaluating the bioactivity of chemical compounds"
  • RF Patent for invention № 2464314 "The method of plasmid DNA condensation for transfection of eukaryotic cells"
  • RF patent for invention № 2468089 "The method of inhibiting the Chlamydia infection pathogen using Lachesanatarabaevi spider genes coding antimicrobial peptides"

 

Technical facilities

Technical facilities

Cultivation of microorganisms proceeded in BioFlo 110 fermentor (New Brunswick Scientifist, USA).

Purification of the recombinant proteins is carried out at the modern liquid chromatograph AKTA FPLC (Amersham).

The laboratory is also fitted with all necessary equipment for genetic engineering and working with cell cultures (confocal microscope Nikon Eclipse E800, laminar flow cabinets, incubators, etc.).

Publications

Publications

  1. Anastasia Nikitina, Vladislav Babenko, Tatyana Akopian, Dmitriy Shirokov, Valentin Manuvera, Alexey Kurdyumov, Elena Kostryukova & Vassili Lazarev, Draft mitochondrial genomes of Hirudo medicinalis and Hirudo verbana (Annelida, Hirudinea), Journal Mitochondrial DNA Part B Volume 1, 2016 - Issue 1
  2. T. N. Safonova, N. N. Mordkovich, V. P. Veiko, N. A. Okorokova, V. A. Manuvera, P. V. Dorovatovskii, V. O. Popov and K. M. Polyakov, Concerted action of two subunits of the functional dimer of Shewanella oneidensis MR-1 uridine phosphorylase derived from a comparison of the C212S mutant and the wild-type enzyme, Acta Cryst. (2016). D72, 203-210 doi:10.1107/S2059798315024353
  3. P. Bobrovsky, V. Manuvera, N. Polina, O. Podgorny, K. Prusakov, V. Govorun, V. Lazarev, Recombinant Human Peptidoglycan Recognition Proteins Reveal Antichlamydial Activity, Infection and immunity, 84 (2016) 2124-2130.
  4. Evgeny D. Nekrasov, Vladimir A. Vigont, Sergey A. Klyushnikov, Olga S. Lebedeva, Ekaterina M. Vassina, Alexandra N. Bogomazova, Ilya V. Chestkov, Tatiana A. Semashko, Elena Kiseleva, Lyubov A. Suldina, Pavel A. Bobrovsky, Olga A. Zimina, Maria A. Ryazantseva, Anton Yu. Skopin, Sergey N. Illarioshkin, Elena V. Kaznacheyeva, Maria A. Lagarkova and Sergey L. Kiselev, Manifestation of Huntington’s disease pathology in human induced pluripotent stem cell-derived neurons, Molecular Neurodegeneration201611:27 DOI: 10.1186/s13024-016-0092-5
  5. Podgorny OV, Lazarev VN, Laser microdissection: A promising tool for exploring microorganisms and their interactions with host, J Microbiol Methods. 2016 Jan 13. pii: S0167-7012(16)30001-X. doi: 10.1016/j.mimet.2016.01.001.
  6. Alexey S. Kurdyumov, Valentin A. Manuvera, Isolda P. Baskova and Vassili N. Lazarev, A comparison of the enzymatic properties of three recombinant isoforms of thrombolytic and antibacterial protein—Destabilase-Lysozyme from medicinal leech, BMC Biochemistry201516:27 DOI: 10.1186/s12858-015-0056-3
  7. Livshits MA, Khomyakova E, Evtushenko EG, Lazarev VN, Kulemin NA, Semina SE, Generozov EV, Govorun VM, Isolation of exosomes by differential centrifugation: Theoretical analysis of a commonly used protocol, Sci Rep. 2015 Nov 30;5:17319. doi: 10.1038/srep17319.
  8. Lobas AA, Karpov DS, Kopylov AT, Solovyeva EM, Ivanov MV, Ilina IY, Lazarev VN, Kuznetsova KG, Ilgisonis EV, Zgoda VG, Gorshkov MV, Moshkovskii SA, Exome-based proteogenomics of HEK-293 human cell line: Coding genomic variants identified at the level of shotgun proteome, Proteomics. 2016 Jul;16(14):1980-91. doi: 10.1002/pmic.201500349.
  9. Natalia Kalinina, Daria Kharlampieva, Marina Loguinova, Ivan Butenko, Olga Pobeguts, Anastasia Efimenko, Luidmila Ageeva, George Sharonov, Dmitry Ischenko, Dmitry Alekseev, Olga Grigorieva, Veronika Sysoeva, Ksenia Rubina, Vassiliy Lazarev and Vadim Govorun, Characterization of secretomes provides evidence for adipose-derived mesenchymal stromal cells subtypes, Stem Cell Research & Therapy20156:221 DOI: 10.1186/s13287-015-0209-8
  10. D.D. Kharlampieva, V.A. Manuvera, O.V. Podgorny, S.I. Kovalchuk, O.V. Pobeguts, I.A. Altukhov, D.G. Alexeev, V.N. Lazarev, V.M. Govorun. Purification and characterisation of recombinant Bacteroides fragilis toxin-2. // Biochimie (2013). V.95. P.2123-2131.
  11. Lazarev V.N., Shkarupeta M.M., Polina N.F., Kostrjukova E.S., Vassilevski A.A., Kozlov S.A., Grishin E.V., Govorun V.M. Antimicrobial peptide from spider venom inhibits Chlamydia trachomatis infection at an early stage. // Arch Microbiol. (2013). V.195. P.173-179.
  12. Nadezhda F. Polina, Marina M. Shkarupeta, Anna S. Popenko, Alexander A. Vassilevski, Sergey A. Kozlov, Eugene V. Grishin, Vassili N. Lazarev and Vadim M. Govorun. Cyto-Insectotoxin 1a from Lachesana tarabaevi Spider Venom Inhibits Chlamydia trachomatis Infection. // Probiotics & Antimicro. Prot. (2012) V.4. P.208-216.
  13. Подгорный О.В., Лазарев В.Н., Говорун В.М. Лазерная микродиссекция в биологии и медицине. // Обзор. Цитология. (2012). Т 54. №5. стр. 381-389.
  14. Alexeev D., Kostrjukova E., Aliper A., Popenko A., Bazaleev N., Tyakht A., Selezneva O., Akopian T., Prichodko E., Kondratov I., Chukin M., Demina I., Galyamina M., Kamashev D., Vanyushkina A., Ladygina V., Levitskii S., Lazarev V., Govorun V. Application of Spiroplasma melliferum Proteogenomic Profiling for the Discovery of Virulence Factors and Pathogenicity Mechanisms in Host-associated Spiroplasmas. // J Proteome Res. (2012). V.11. P. 224-236.
  15. Vishnyakov I.E., Levitskii S.A., Manuvera V.A., Lazarev V.N., Ayala J.A., Ivanov V.A., Snigirevskaya E.S., Komissarchik Y.Y., Borchsenius S.N. The identification and characterization of IbpA, a novel α-crystallin-type heat shock protein from mycoplasma. // Cell Stress Chaperones. (2012). V.17. P.171-180.
  16. Lazarev V.N., Polina N.F., Shkarupeta M.M., Kostrjukova E.S., Vassilevski A.A., Kozlov S.A., Grishin E.V., Govorun V.M. Spider Venom Peptides for Gene Therapy of Chlamydia Infection. // Antimicrob Agents Chemother. (2011). V.55. P. 5367–5369.
  17. Lazarev V.N., Levitskii S.A., Basovskii Y.I., Chukin M.M., Akopian T.A., Vereshchagin V.V., Kostrjukova E.S., Kovaleva G.Y., Kazanov M.D., Malko D.B., Vitreschak A.G., Sernova N.V., Gelfand M.S., Demina I.A., Serebryakova M.V., Galyamina M.A., Vtyurin N.N., Rogov S.I., Alexeev D.G., Ladygina V.G., Govorun V.M. Complete genome and proteome of Acholeplasma laidlawii. // J Bacteriol. (2011). V.193. P. 4943-4953.
  18. Полина Н.Ф., Лазарев В.Н. Генетическая терапия инфекционных заболеваний. Генотерапевтические агенты и невирусные системы доставки генетического материала. // Обзор. Эфферентная и физико-химическая медицина (2011). №1. стр. 11-20.
  19. Levitskiy S.A., Sycheva A.M., Kharlampieva D.D., Oberto J., Kamashev D.E., Serebryakova
    M.V., Moshkovskii S.A., Lazarev V.N., Govorun V.M. Purification and functional analysis of recombinant Acholeplasma laidlawii histone-like HU protein. // Biochimie (2011). V.93. P.1102-1109.
  20. Lazarev V.N., Borisenko G.G., Shkarupeta M.M., Demina I.A., Serebryakova M.V., Galyamina M.A., Levitskiy S.A., Govorun V.M. The role of intracellular glutathione in the progression of Chlamydia trachomatis infection. // Free Radic Biol Med. (2010). V.49. P.1947-1955.