Recombinant plasmid dna purvgf, which determines the synthesis of the growth factor, vaccinia virus strain l-ivp together with beta - galactosidase of escherichia coli, and bacterial strain escherichia coli containing a recombinant plasmid dna purvgf - producer of the growth factor, vaccinia virus strain l-ivp

 

(57) Abstract:

Usage: biotechnology, drug development, causing cell proliferation, which can be used for wound healing. Essence: construction of recombinant plasmid DNA, which determines the synthesis of the growth factor, vaccinia virus (FRWO), and obtaining strain containing this plasmid. The strain used as a producer of FRWO, fused with beta-galactosidase. Recombinant FRO in the long term can be applied as a mitogen cells of epithelial origin. 2 C. p. F.-ly, 3 ill.

The invention relates to biotechnology and medicine, in particular to biologically active agents and methods for their preparation methods of genetic engineering.

As it became known, the growth factor, vaccinia virus (VGF) related to the family of EGF-like factors [1] Mature molecule VGF has 35% amino acid homology with epidermal growth factor (EGF) and binds to the EGF receptor on the cell surface, causing cell proliferation [2]

Under normal conditions VGF is produced by vaccinia virus in his reproductions in cultures of cells, and the amount of this protein in the culture medium is very small,P> Creating bacterial producer VGF will get it in preparative quantities and purified to a homogeneous state, using a relatively simple method of purification.

The literature describes the VGF gene of strain WR-vaccinia virus, as well as the positive impact of this factor, isolated from the culture medium of cells infected with vaccinia virus, or chemically synthesized, on cell proliferation in vitro [4,6]

Also known gene that encodes the structure of Hind III G-fragment DNA vaccine strain L-IVP of vaccinia virus growth factor, vaccinia virus [3,5] VGF Gene isolated from clonal variant (clone 4) strain L-IVP of vaccinia virus differs from the VGF gene of strain WR described in the literature [4] four nucleotide substitutions at positions 86, 319, 358, 359. which leads to the replacement of the amino acid in position 29 of serine to leucine, at position 107 of methionine for valine and at position 120 of leucine for serine [5 prototype]

However, information regarding the creation of recombinant producers VGF suitable for producing this protein in preparative quantities not available.

The aim of the invention is the creation of a recombinant strain of E. coli bacteria-producer VGF opvattingen the United States is based plasmid DNA pUR VGF, coding the synthesis of VGF, which has a size of 5,419 N. p. and consists of the following elements (Fig. 1):

vector part in the form of a Hind III BamH I fragment of plasmid pUR 290 size 5,186 as N. p. containing the LacZ gene with its own promoter bla-gene.

BamHI Hind III fragment size 233 N. p. contains a sequence of open frame broadcast (ORT) VGF gene of vaccinia virus strain L-IVP without sequences encoding signal and transmembrane part of the VGF gene (Fig. 3)

Plasmid DNA pUR VGF contains unique restriction sites for the enzymes BamHI, Hind III, Tag I, Ava II, Alu I, Nla III, Cla I.

To achieve this purpose a method of constructing plasmids pUR VGF, namely, that the vector part, obtained by cleavage of plasmid DNA pUR 290 restriction endonucleases BamH I, Hind III, merge with DNA ligase of phage T4 with a DNA fragment of vaccinia virus treated with the same endonucleases, which was pre-amplified by polymerase chain reaction using pre-synthesized primers complementary terminal sequences of the DNA of vaccinia virus coding for Mature VGF. Additionally, at the beginning of the gene VGF enter the website, coding for the mH I.

A significant advantage of the proposed method is that it achieves expression virousspecificakih protein VGF in E. coli cells. Gene protein VGF is in the same reading frame with the LacZ gene, which provides a synthesis of the chimeric polypeptide mol. m 126 kDa, consisting of beta-galactosidase and protein VGF, United cyclotouring connection.

Plasmid pUR VGF not described in literature and for the first time obtained in this work.

To obtain the producer strain E. coli competent cells of strain WMN transformed with recombinant plasmid DNA. Producing strains WMN differs from the original only by signs, the transferred plasmid, i.e. is resistant to ampicillin and synthesizes chimeric protein CD consisting of beta-galactosidase of E. coli and protein VGF and with beta-galactosidase activity. Output VGF is 6 mg/g of bacterial cells. The obtained bacterial strain producing registered in the all-Union collection of industrial microorganisms Institute of genetics under number CPSU/M-6566.

The resulting strain is characterized by the following features.

1. Morphological features

Cells are straight, rod-shaped form, 1,2x1,6x2,0-6,0 μm, the Ki

Cells grow well on simple nutrient media, optimum growth temperature is 30-37oC, pH of 6.8 to 7.5.

With growth mesopatamia and fish agar develop in the form of smooth, round, shiny, grey, muddy colonies with smooth edges. When operating in liquid environments cause uniform turbidity from sediment.

3. Physiological and biological characteristics

The optimal cultivation temperature of 37oC, the optimum pH of 6.8 to 7.5. As the carbon source used by many carbohydrates, alcohols, organic acids. As the source of nitrogen used as mineral salts or ammonium nitrate and organic compounds, peptone, amino acids.

4. Antibiotic resistance

Resistant to ampicillin due to the presence of plasmids pUR VGF at a concentration of 70 to 100 mg/L.

5. The stability of the plasmid in the strain

While maintaining cell strain for 1.5 years on fish agar at +40oC or 30% glycerol at -70oC in the presence of ampicillin is not observed rearrangement or loss of the plasmid DNA.

The invention consists in that the cloning of the gene VGF virus ospovat the life of the chimeric polypeptide 126 kDa (beta-galactosidase-VGF), positively reacting with monospecific serum to a synthesized peptide corresponding 20-33 amino acid residues N-Terminus of the Mature molecule VGF (Fig. 2).

In Fig. 1 shows a physical map of plasmid pUR VGF; Fig. 2 - immunoblotting of cell lysate of E. coli strain UMNS 7118 after a three-hour induction IPTG (isopropyl-thio-beta-D galactopyranoside) with monospecific anticorodal rabbit received by the synthesized peptide (amino acid residues 20-33 N-Terminus of the Mature VGF).

Tracks:

1 proteins of E. coli cells transformed with plasmid pUR VGF, separated by electrophoresis in 8% SDS page;

2 proteins cells pUR 290 transformed by the plasmid pUR 290, separated by electrophoresis in 8% SDS page.

In Fig. 3 shows the nucleotide sequence encoding a Mature VGF.

Example 1. Construction of recombinant plasmid DNA pUR VGF.

Using synthetic oligonucleotide primers for polymerase chain reaction (PCR) and viral DNA (strain L-IVP), get a DNA fragment encoding Mature VGF. Options full-size chain reaction following: annealing - 37o, 1 min; elongation 72o, 1 min; denaturation 95o, 1 min; the number of cycles 25; enzyme-T is ogeny sites for restricted BamHI and HindIII, as well as a stop codon.

After amplification gain 5 μg of the DNA fragment mobility in SDS page 230 p. N. the Specified DNA fragment is treated with enzymes BamHI and HindIII under standard conditions (50 mm NaCl, mm mercaptoethanol, 10mm MgCl2, 10mm Tris HCl, pH 7.5); the reaction is stopped by adding 0.5 m EDTA (5 µl per 100 µl of the mixture), remove restrictase phenol from the aqueous phase precipitated DNA, as described in[7]

The vector is prepared as follows: 5 μg DNA pUR 290 cut in standard conditions BamHI and HindIII by restrictase according to the method described in [7] and put 5% SDS page. Allocate bandwidth DNA and dissolved in 50 μl TE.

Vector DNA (0.05 mg) and the fragment (1 μg) sew with T4-DNA-ligase [7] Recombinant clones are selected restriction fragments length polymorphism analysis. Competent cells prepared from E. coli strain JM109.

Example 2. The receipt of the producer strain E. coli BMH 7118.

Bacterial cells of E. coli JM109 containing plasmid DNA pUR VGF grown in 4 ml of L-broth with ampicillin at a concentration of 40 μg/ml to titer h cells/ml Plasmid DNA secrete the alkaline method [3] and retransforming in cells of E. coli strain BMH 7118. The resulting strain is subjected to analysis for the presence of plasmid markers.

1. Resistance to ampicillin.

2. Expression of the chimeric polypeptide 126 kDa.

Cells of E. coli strain WMN induce IPT G (IOE-3 M) and are lysed in 3% LTOs (50 mm 2-mercaptoethanol; 20 mm Tris-HCl pH 6.8; 30% sucrose 5 ml) at 95oC. the Denatured polypeptides share the disk-electrophoresis in 8% SDS page and detected using Kumasi R-250. The molecular weight of the chimeric polypeptide 126 kDa calculated from the electrophoretic mobility.

3. Permentally activity of the chimeric polypeptide 126 kDa.

Cells of strain is plated on agar medium containing 40 μg/ml of ampicillin, 0.1 mg/ml X-gal, 0.1 ág/ml IPT, Colonies of E. coli strain JM109 have a blue color.

4. The formation of AG-AB complex chimeric protein of 126 kDa with monospecific anticorodal to the synthesized peptide (amino acid residues 20-33 of the Mature molecule VGF) immunogenic properties of the chimeric protein of 126 kDa confirmed in response immunoblotting (Fig.2), as described below.

Example 3. Immunochemical analysis of the chimeric polypeptide 126 kDa.

Bacterial strain E. coli BMH pUR VGF pokasivaut to D 0,6 O. E. and after adding IPT G to 50 mg/ml and grown for 3 h at 37oC, active mixing. 1 ml cell culture centrifuged 3 min at 6000 sediment suspending solution separated in 8% SDS page and carry out the transfer of polypeptides to nitrocellulose. Fixed protein preparation incubated in 1% BSA solution, then at room temperature, incubated with monospecific rabbit anticorodal against synthesized peptide 1 h 150 mm NaCl; 20 mm Tris-HCl, pH 7,7; 0.2% Triton X-100. Washed three times in buffer of composition and incubated for 1 h with peroxidase-conjugated protein A. After washing in buffer, dye AG-AB complex with 3,3-diaminobenzidine HCl.

The invention allows to obtain VGF in milligramme quantities and to explore its immunological and biological properties.

Received the producer of recombinant VGF can be used to study the interaction of the growth factor membrane receptors, obtaining antibodies to VGF and in the future possible use of recombinant VGF obtained in E. coli cells as a mitogen, causing the proliferation of cells of epithelial origin in vivo and in vitro.

Thus, the results of studies obtained producing strains of E. coli expressing the growth factor, vaccinia virus. Further studies of this protein will allow you to create a database for use in the treatment of burn disease and for healing difficult shivaami trophic lang is 1987, V. 235, N4786, P. 80-82.

Petrov, C. S. Khromykh, A. A. Dmitriev, I. P. and other Genetic and cell engineering in the solution of fundamental problems of biotechnology: proceedings of all-Union Conf. Tartu, 1989, so I, c. 171-174.

Venkatesan S. Gershowitz A. Moss B. J. of Virology, 1982, V. 44, p. 637-646.

Petrov Century North Cheshenko N. In. Belavin P. A. and other Biotechnology, 1992, N5, S. 35-39.

Lin J. Caporaso C. Ke, X. H. Tam.J. of Biol. Chemistry, 1990,V. 265, 31 N., P. 1881-1890.

Maniatis T. Fritsch H., Sambrook J. Methods of genetic engineering. Molecular cloning. TRANS. from English. M. Mir, 1984.

Salganik R. I. Methods of molecular genetics and genetic engineering. Novosibirsk, "Nauka" 1990.

1. Recombinant plasmid DNA pURVGF, which determines the synthesis of the growth factor, vaccinia virus strain L-IVP together with beta-galactosidase of Escherichia coli, size 5419 p. O. containing Bam HI Hind III DNA fragment from plasmid pUR 290 size 5186 p. O. corresponding gene of resistance to ampicillin and gene beta-galactosidase, Bam HI Hind III fragment of the genome of vaccinia virus size 233 p. O. including gene growth factor vaccinia virus encoding the Mature polypeptide, one cleavage site for restrictase Bam HI, one cleavage site for restrictase Tag I, located at a distance of 17 p. O. from the Bam HI site is ment to restrictase Ava II, located at a distance of 79 p. O. from the Bam HI site, two sites of cleavage for restrictase Nla III, located at a distance of 160 p. O. and 187 p. O. from the Bam HI site, a single site of cleavage for restrictase Hind III, located at a distance of 245 p. O. from the Bam HI site, a single site of cleavage for restrictase Cla I, located at a distance of 246 p. O. from the Bam HI site, a genetic marker: a gene beta-lactamase, providing resistance to ampicillin.

2. The bacterial strain Escherichia coli VKPM In 6566 containing recombinant plasmid DNA pURV6 F producer growth factor, vaccinia virus strain L-IVP.

 

Same patents:

FIELD: genetic engineering, virology, pharmacy.

SUBSTANCE: invention proposes the recombinant modified virus OF VACCINE Ankara able to express structural antigens of hepatitis C virus. Virus comprises DNA sequences encoding structural antigens of hepatitis C virus or their functional regions or epitopes of hepatitis C virus structural antigens. Also, invention proposes a pharmaceutical composition comprising such virus, eucaryotic cell infected with such virus, a method for preparing such virus and a method for preparing hepatitis C virus structural polypeptides. Invention can be used in virology and medicine for preparing hepatitis C virus antigen.

EFFECT: valuable properties of virus.

20 cl, 14 dwg, 1 tbl

FIELD: biotechnology, virology, medicine.

SUBSTANCE: invention relates to attenuated virus derived from modified Ankara vaccina virus. Said virus are not able for reproduction by replication in human cell lines. Also disclosed are application of virus or recombinant variants thereof as drug or vaccine, as well as method for inducing of immune response in patients with defected immunity, in patients having immunity to vaccine virus, or in patient during antiviral therapy.

EFFECT: variant of Ankara vaccina virus effective in medicine and veterinary.

86 cl, 15 dwg, 1 tbl, 2 ex

FIELD: biotechnology, protein engineering.

SUBSTANCE: claimed library represents E.coli TGI cells wherein each cell contains fragmid DNA providing biosynthesis of filamentous bacteriophages exposing unique human single-stranded antibody on surface thereof. Also disclosed is recombinant fragmid pHEN-2A8 DNA containing artificial gene of human single-stranded antibody under control of lactose operon promoter providing synthesis of human single-stranded antibody in composition of chimerical protein with membrane pIII protein of M13 bacteriophage in E.coli cells. Also disclosed is method for production of artificial human single-stranded 2A8 antibody by using such fragmid DNA.

EFFECT: fragmid library useful in medicine.

3 cl, 7 dwg, 10 ex

FIELD: biotechnology, in particular gene engineering.

SUBSTANCE: Gene of B9R protein having high homology with extracellular segment of interferon-gamma receptor is isolated by PCR method from Mankeypox virus genome of strain Zaire-96-1-16. Then said protein is cloned in donor plasmid pFastbAC and via site-specific transposition recombinant bacmid is constructed. Said bacmid is used for pest cell transfection to generate target strain.

EFFECT: new drugs for treatment of human diseases associated with hyperproduction of interferon-gamma.

2 cl, 3 dwg, 6 ex

FIELD: molecular biology, gene engineering, medicine.

SUBSTANCE: disclosed is method for determination of different orthopoxviruses based on one-step PCR followed by hybridization of obtained fluorescence labeled amplicones with DNA microarray containing discriminating orthopox- and herpesvirus-hybridization probes.

EFFECT: method for express-diagnosis of orthopoxviruses and discrimination thereof from herpesviruses.

3 cl, 14 dwg, 3 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: invention is related to preparation of protein, binding tumour necrosis factor (TNF), and may be used in medicine. Strain-producer of baculovirus BvG2RIgG is created with the help of recombinant plasmid DNA pFastBac-G2R-IgG with size of 6444 p.n. and molecular mass 4.18 mDa, which bears fragment of smallpox virus genome of strain India-1967, which codes protein that binds TNF, and fragment of human genome, which codes fragment of heavy chain of human antibody G. Produced strain produces soluble chimeric protein, which consists of smallpoz virus protein, which binds TNF, and fragment of heavy chain of human antibody G.

EFFECT: wider spectrum of new generation preparations intended for treatment of human diseases related to hyperproduction of tumour necrosis factor.

2 cl, 3 dwg, 1 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: set contains species-specific oligonucleotide primer pairs and appropriate fluorescent-marked probes for conducting one-stage instant identification of several human-pathogenic Orthopoxviruses (VARV, MPXV, CPXV and VACV) by means of real-time multiplex PCR.

EFFECT: invention is intended for instant diagnostics of human and animal Orthopoxvirus infections by real-time multiplex PCR.

10 dwg, 2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: there are presented two recombinant plasmid DNA pFastBac 1 -G2R-dSECRET and pQE-60-TNFR-CrmB-Ind-67 coding TNF-binding CrmB protein domain. Said recombinant plasmid DNA pQE-60-TNFR-CrmB-Ind-67 is designed for transformation in cells of the strain E.coli SG13009[pRep4] - a producer of TNF-binding CrmB BHO protein domain.

EFFECT: presented group of inventions is applicable for preparing drugs used in therapy of severe human diseases caused by hyperproduction of tumour necrosis factor and may be used in medicine.

3 cl, 3 dwg, 12 ex

FIELD: biotechnologies.

SUBSTANCE: inventions represent synthetic oligonucleotide primers: P1 - 5'-TGGTTACTATTCCATCACCATT-3' (annealing site 11-32 base pairs), P2 - 5'-CGAAACGTCACTTTCGCAAC-3' (annealing site 259-278 base pairs) and method for detection of DNA of infectious anaemia virus of chickens with their help. The method consists in the fact that primers flank the section of the virus genome, which includes CpG islands and VNTR repetitions in the polymerase chain reaction in real time mode. In case of positive reaction, a peak of the melting curve 92C° appears, and when the reaction is confirmed by means of electrophoresis a fragment is visualised in the gel that corresponds to the size of 268 base pairs.

EFFECT: method of diagnostics makes it possible to determine quantitative content of a virus in tissues and may be used to diagnose infectitious anaemia of chickens.

2 cl, 3 dwg, 4 tbl, 5 ex

FIELD: biotechnology.

SUBSTANCE: recombinant plasmid DNA pGEM-Puro-DS-Apo is proposed comprising synthetic apoptin gene flanked by sequences of vaccinia virus genome of island C10L-C12L, and the recombinant strain VVdGF-ApoS24/2 of vaccinia virus producing apoptin.

EFFECT: invention can be used to develop medicinal products to control cancerous diseases.

2 cl, 5 dwg, 1 tbl, 6 ex

FIELD: biotechnology, veterinary science.

SUBSTANCE: invention relates to therapeutic vector used in therapy of infectious diseases in cats that comprises at least one foreign nucleic acid each of that (a) encodes protein taken among the group consisting of feline protein CD28 represented in SEQ ID NO:8 or its immunogenic moiety; feline protein CD80 represented in SEQ ID NO:2 or 3, or its immunogenic moiety; feline protein CD86 represented in SEQ ID NO:6 or its immunogenic moiety, or feline protein CTLA-4 represented in SEQ ID NO:10 or its immunogenic moiety; and (b) nucleic acid that is able to be expressed in insertion of vector in the corresponding host. Indicated therapeutic vector is used in effective dose as component of vaccine against infectious diseases in cats for their immunization and in methods for enhancement or inhibition of immune response in cats and reducing or eradication of tumor in cats. Invention provides stimulating the activation and proliferation of T cells and to enhance effectiveness of control of infectious diseases in cats.

EFFECT: valuable biological properties of recombinant virus.

41 cl, 13 dwg

FIELD: gene engineering, in particular apoptosis inducing gene delivery vectors useful for cancer, hyperplasia, metaplasia and displasia diagnosis and treatment.

SUBSTANCE: recombinant adenovirus apoptin-containing vectors are obtained by cotransfection into 911 helper cell line of p.Amb-VP3 adaptor plasmids (in case of VP3 protein expression) or pMAb-VP2 plasmids (in case of VP2 protein expression) and JM17 DNA. p.Amb-VP3 plasmids carry apoptin gene in 5'-3'-orientation, expressing under control of adenoviral main late promoter. Plasmid JM17 DNA contains complete adenoviral DNA excepted E1 and E2 regions. pMAb-°VP2 plasmids carry apoptin gene with two point mutation in limits of coding region. Cotransfections are carried out by calcium phosphate method. Recombinant adenoviral DNA is formed by homologous recombination between homologous viral sequences representing in p.Amb-VP3 (or pMAb-VP2) plasmid and in adenoviral DNA from plasmid JM17 DNA. Cell infection of various human tumors with gene delivery vectors causes to tumor cell apoptosis induction and sufficiently reduced normal, diploid, non-transformed or non-pernicious cell apoptosis.

EFFECT: new gene delivery vector capable to induce cell apoptosis.

8cl, 7 dwg

FIELD: gene engineering, in particular apoptosis inducing gene delivery vectors useful for cancer, hyperplasia, metaplasia and displasia diagnosis and treatment.

SUBSTANCE: recombinant adenovirus apoptin-containing vectors are obtained by cotransfection into 911 helper cell line of p.Amb-VP3 adaptor plasmids (in case of VP3 protein expression) or pMAb-VP2 plasmids (in case of VP2 protein expression) and JM17 DNA. p.Amb-VP3 plasmids carry apoptin gene in 5'-3'-orientation, expressing under control of adenoviral main late promoter. Plasmid JM17 DNA contains complete adenoviral DNA excepted E1 and E2 regions. pMAb-°VP2 plasmids carry apoptin gene with two point mutation in limits of coding region. Cotransfections are carried out by calcium phosphate method. Recombinant adenoviral DNA is formed by homologous recombination between homologous viral sequences representing in p.Amb-VP3 (or pMAb-VP2) plasmid and in adenoviral DNA from plasmid JM17 DNA. Cell infection of various human tumors with gene delivery vectors causes to tumor cell apoptosis induction and sufficiently reduced normal, diploid, non-transformed or non-pernicious cell apoptosis.

EFFECT: new gene delivery vector capable to induce cell apoptosis.

8cl, 7 dwg

FIELD: gene engineering, in particular apoptosis inducing gene delivery vectors useful for cancer, hyperplasia, metaplasia and displasia diagnosis and treatment.

SUBSTANCE: recombinant adenovirus apoptin-containing vectors are obtained by cotransfection into 911 helper cell line of p.Amb-VP3 adaptor plasmids (in case of VP3 protein expression) or pMAb-VP2 plasmids (in case of VP2 protein expression) and JM17 DNA. p.Amb-VP3 plasmids carry apoptin gene in 5'-3'-orientation, expressing under control of adenoviral main late promoter. Plasmid JM17 DNA contains complete adenoviral DNA excepted E1 and E2 regions. pMAb-°VP2 plasmids carry apoptin gene with two point mutation in limits of coding region. Cotransfections are carried out by calcium phosphate method. Recombinant adenoviral DNA is formed by homologous recombination between homologous viral sequences representing in p.Amb-VP3 (or pMAb-VP2) plasmid and in adenoviral DNA from plasmid JM17 DNA. Cell infection of various human tumors with gene delivery vectors causes to tumor cell apoptosis induction and sufficiently reduced normal, diploid, non-transformed or non-pernicious cell apoptosis.

EFFECT: new gene delivery vector capable to induce cell apoptosis.

8cl, 7 dwg

FIELD: genetic engineering, proteins, medicine, pharmacy.

SUBSTANCE: invention relates to a method for preparing a fused protein representing immunoglobulin Fc-fragment and interferon-alpha and can be used in treatment of hepatitis. Method involves construction of a fused protein comprising immunoglobulin Fc-fragment prepared from Ig G1 or Ig G3 in direction from N-end to C-end and the end protein comprising at least one interferon-alpha. Fc-fragment and the end protein are joined directly or by a polypeptide bridge. The fused protein is used for preparing a pharmaceutical composition used in treatment of liver diseases and in a method for targeting interferon-alpha into liver tissues. Invention provides preparing the fused protein eliciting with biological activity of interferon-alpha providing its concentrating in liver and showing enhanced solubility, prolonged half-time life in serum blood and enhanced binding with specific receptors.

EFFECT: improved targeting method, valuable biological properties of fused protein.

10 cl, 5 dwg, 9 ex

FIELD: genetic engineering, proteins, medicine, pharmacy.

SUBSTANCE: invention relates to a method for preparing a fused protein representing immunoglobulin Fc-fragment and interferon-alpha and can be used in treatment of hepatitis. Method involves construction of a fused protein comprising immunoglobulin Fc-fragment prepared from Ig G1 or Ig G3 in direction from N-end to C-end and the end protein comprising at least one interferon-alpha. Fc-fragment and the end protein are joined directly or by a polypeptide bridge. The fused protein is used for preparing a pharmaceutical composition used in treatment of liver diseases and in a method for targeting interferon-alpha into liver tissues. Invention provides preparing the fused protein eliciting with biological activity of interferon-alpha providing its concentrating in liver and showing enhanced solubility, prolonged half-time life in serum blood and enhanced binding with specific receptors.

EFFECT: improved targeting method, valuable biological properties of fused protein.

10 cl, 5 dwg, 9 ex

FIELD: biotechnology, medicine, in particular viral disease treatment.

SUBSTANCE: invention relates to recessive dividing retroviral vector useful in inhibition of wild-type retrovirus replication. Vector contains retroviral long terminal repeat sequences; retroviral packing signal; nucleotide sequence encoding (expressing) genetic antiviral agent; and optionally the second nucleotide sequence. Disclosed are method for production of said vector and reproduction thereof. Further isolated and purified nucleic acid (NA) molecule providing of selective advantage in regard to viral generation packing into virions is disclosed. Uses of retroviral vector in particular for specific antibody production are described.

EFFECT: new genetic antiviral agents generating prolonged and stable immunological responses in regard, for example, to AIDS and cancer viruses.

97 cl, 11 ex

FIELD: biotechnology, medicine, in particular viral disease treatment.

SUBSTANCE: invention relates to recessive dividing retroviral vector useful in inhibition of wild-type retrovirus replication. Vector contains retroviral long terminal repeat sequences; retroviral packing signal; nucleotide sequence encoding (expressing) genetic antiviral agent; and optionally the second nucleotide sequence. Disclosed are method for production of said vector and reproduction thereof. Further isolated and purified nucleic acid (NA) molecule providing of selective advantage in regard to viral generation packing into virions is disclosed. Uses of retroviral vector in particular for specific antibody production are described.

EFFECT: new genetic antiviral agents generating prolonged and stable immunological responses in regard, for example, to AIDS and cancer viruses.

97 cl, 11 ex

FIELD: biotechnology, virology, medicine.

SUBSTANCE: invention relates to attenuated virus derived from modified Ankara vaccina virus. Said virus are not able for reproduction by replication in human cell lines. Also disclosed are application of virus or recombinant variants thereof as drug or vaccine, as well as method for inducing of immune response in patients with defected immunity, in patients having immunity to vaccine virus, or in patient during antiviral therapy.

EFFECT: variant of Ankara vaccina virus effective in medicine and veterinary.

86 cl, 15 dwg, 1 tbl, 2 ex

FIELD: gene engineering.

SUBSTANCE: the present innovation deals with the ways for obtaining transgenic poultry due to introducing retroviral vectors into blastodermal cells through the fissure in the shell of nonhatching egg from the side of its blunt end. With the help of insulin syringe one should introduce gene constructions for the depth of about 2-3 cm near a germinal disk. The innovation enables to simplify the procedure of introducing gene constructions into target cells at maintaining general efficiency of transgenesis that leads to the decrease of embryonic lethality.

EFFECT: higher efficiency.

Up!