>>

a problem Urgency


Agrobacterium tumefaciens is the eurysynusic kind of soil bacteria invoking formation koronchatyh of Gauls at plants which result from fragment embedding Ti - plasmids (T-DNK) of agrobacteria in a vegetative genome and expressions of bacterial genes in a cage-recipient.
One of key factors in the course of transmission T-DNK is bacterial protein VirE2 which co-operates with otsdnk and forms a transported complex (T - a complex). By means of electronic microscopy it is shown, that VirE2 forms with otsdnk the T-complex with the spiral frame well approaching for problems of protection by DNA and nuclear import [Frenkiel-Krispin et al., 2006]. Now it is established, that protein carries out some functions: linkage with T-ДНК, its protection against degradation vegetative endonukleazami, transport T-DNK through vegetative membranes. It is besides, shown, that VirE2 co-operates with vegetative cytoplasmatic squirrels-shaperonami (Roca, Roc4 and CypA) and protein VIP2 which provide interaction with a chromatin and promote integration T-DNK [Tzfira, Citovsky, 2002]. Transport T-DNK in a cage of the master, possibly, can be realised by different pathes. Formation of a pore from agrobacterial protein VirE2 in lipidnoj to a membrane can be one of them. Thus there are data, that VirE2 is capable to be built in an artificial membrane and to form in it the canal which is discovered at imposing of electric field with intensity 10-100 mv [Dumas et al., 2001; CHumakov with soavt., 2010]. Another possible by penetration T-DNK into zooblasts can be endotsitoz. [See CHumakov, 2001]. Nevertheless, experimental proofs of absorption T-DNK a cage by endotsitoza now
No.
In the modern literature the phenomenon of transmission of short oligonucleotides through artificial membranes and T-DNK agrobacteria through membranes of zooblasts at participation of protein VirE2 is described in works [Kunik et al., 2001; Petrunia et al., 2008]. However, the hit mechanism otsdnk in a cage, mediated by protein VirE2, maloissledovan. The purpose of the given work is analysis of participation of protein VirE2 on delivery otsdnk through a membrane in eukarioticheskuju a cage: through porovyj a complex,
Formed by protein VirE2; with the help endotsitozaz mediated by protein VirE2.
The purposes and research problems

The purpose of the present work was studying of possibility of protein VirE2 and its complexes to participate in transport process otsdnk through membranes eukarioticheskoj cages.
During research following problems were solved: to Construct three-dimensional computer models of complexes on a basis of protein VirE2 and to carry out their analysis on vstrojku in a membrane and presence of canals. To study supramolecular complexes on the basis of VirE2 the squirrel and otsdnk - VirE2 biocomputer science methods. To secure, clear rekombinantnyj squirrels VirE2 and to carry out analysis of its properties in vitro. To study transmission of oligonucleotides to zooblasts at presence rekombinantnogo squirrel VirE2.
Scientific novelty of work:
For the first time analysis of complexes from two and four proteins VirE2 is carried out by a method of normal fashions. It is established, that in some fashions the possible portal mechanism of canals at complexes from two proteins VirE2 is observed.
For the first time complexes from six proteins VirE2 are simulated.
For the first time analysis of protein VirE2 in a complex with protein-shaperonom VirEl methods of molecular dynamics is carried out. It is established, that the model of proteins VirE2-VirE1 is in an equilibrium, stable state at time of modelling to 500 ps. Also it is established, that the frame of protein VirE1 in complex VirE2-VirE1 possesses the greatest motility.
For the first time it is established, that rekombinantnyj protein VirE2 promotes accumulation of short synthetic oligonucleotides in nativnyh cages of line HeLa, but not in cages SPEV.
The scientifically-practical importance of work
As protein VirE2 promotes transmission oлигoнуклеoтидoв in zooblasts, further this phenomenon can be used by working out bezvirusnyh technologies of delivery of genes in zooblasts and technologies genoterapii.
The substantive provisions of the dissertation born on protection of Squirrels VirE2 is capable to form in silico complexes of two, four, six subunits which are capable to be built in a membrane. Models of complexes from protein VirE2 are stable and possess canals, and the complex from two proteins VirE2 has the portal canal. Protein VirE2 promotes hit of oligonucleotides in zooblasts HeLa, but not in cages SPEV, and also co-operates with otsdnk.
The contribution of the competitor to the results received in cooperation:
The contribution of the author of dissertation consists in reception of primary data on modelling of complex frames on a basis of protein VirE2 in silico, to transmission analysis otsdnk in zooblasts, to computer analysis of albuminous complexes from protein VirE2 which are spent together with Mazilovym V. I, Volohinoj I.V. and Chumakovym M. I on base a lab.
Bioengineerings IBFRM the Russian Academy of Sciences. Results are published in joint articles. Analysis of model of protein VirE2 in a complex with protein shaperonom VirE1 methods of molecular dynamics, analysis of complexes from two and four proteins VirE2 methods of normal fashions, and also statistical processing of experimental data are spent independently. Planning of experiments, their interpretation, analysis and preparation of results were spent under the guidance of M.I.Chumakova.
Work is executed on base a lab. Bioengineerings IBFRM the Russian Academy of Sciences in 20112013 within the limits of NIR «Research of transmission of DNA-albuminous of complexes in eukarioticheskie cages», № 01200606182 (nauch. Arms. d.b.n. CHumakov M.). Work is sustained by the Russian Federal Property Fund grant №11-04-01331 «Studying of the mechanism of transmission T-DNK of agrobacteria in generative cages of corn» (20112013, arms. k.b.n. Volohina I. V), the agreement № 8728 FTSP «Scientific and scientific and pedagogical shots of innovative Russia» for 2009-2013. «Working out of technology of delivery of DNA in zooblasts with the help of protein VirE2» (arms. k.b.n. Mazilov S.I.).
Approbation of results
Dissertation stuffs have been presented on H to the All-Russia conference «Биомеханика-2010», (Saratov, on May, 16-22th, 2010), V Vseros. To school-konf. A pier. uchen. «Strategy of interaction of microorganisms and plants with environment» (Saratov, 28 sent.-1 okt. 2010), on XIV International school-conference for a pier. uchen. And stud. On optics, the laser physics and biofotonike «Saratov Fall Meeting 10» (Saratov, on October, 5-8th, 2010), 2nd annual scientific and technical conferences
nanotehn. obshch. Russia “development Prospects in Russia of NBIK-TECHNOLOGIES as basic scientific direction of break to the sixth technological way” (Moscow, on October, 14-15th 2010), on III mezhdunar. A forum on nanotehnologijam “RUSNANOTECH 2010” (Moscow, on November, 1-3st 2010), at the Moscow international conference on computer biology (Intern. Moscow Conf. on Comp. Mol. Biol. MCCMB ’ 11, Moscow, on July, 21-24st 2011), at IV congress of biophysicists of Russia (N.Novgorod, on August, 20-26th, 2012), at 13th forum of young scientists FEBS (YSF 2013) (St.-Petersburg, on July, 3-5rd, 2013) and 38 Congress FEBS (St.-Petersburg, on July, 6-11th, 2013).
Publications
On a dissertation theme 12 printing works, from them 4 articles in the magazines entering into List VAK are published.
Frame and dissertation volume
The dissertation, in volume of 109 pages, has frame: introduction, the head with the literature review, the head with instructions of the used stuffs and research methods, the head with the description of experimental data and their discussion, conclusions, the list of the quoted literature from 142 sources, from which 130 - foreign, appendices. The dissertation contains 44 drawings and 6 tables.
| >>
A source: GUSEV Jury Sergeevich. FRAME And FUNCTIONS of PROTEIN VirE2 In TRANSMISSION otsdnk VEUKARIOTICHESSKY CAGES. 2014

More on topic a problem Urgency:

  1. a problem Urgency.
  2. a problem Urgency
  3. a problem Urgency
  4. the Urgency of a problem of research
  5. a problem Urgency
  6. the problem Urgency
  7. a problem Urgency.
  8. a problem Urgency
  9. the Urgency of a problem of research
  10. the Urgency of a problem of research
  11. the Urgency of a problem of research
  12. the Urgency of a problem of research
  13. the Urgency of a problem of research
  14. the Urgency of research and problem statement
  15. the Urgency of a problem and some risk factors
  16. a problem Urgency mikoplazmennoj infections in obstetrics and gynecology