Avirulent strain of bacteria vibrio cholerae biovar eltor, serovar ogava (ctxa-, tcpa-, toxr-, zot-), used in immunological and genetic investigations and educatory process

FIELD: biotechnology, in particular immunological and genetic investigations and educatory process.

SUBSTANCE: Claimed avirulent strain of bacteria Vibrio cholerae biovar eltor, serovar Ogava (ctxA-, tcpA-, toxR-, zot-) is isolated from natural water source of open basin in Turkmenistan, doesn't contain ctxA, tcpA, toxR, zot genes and has stable avirulent atoxigenic properties for long-time storage.

EFFECT: strain useful in screening of new drugs for cholera immunoprophylaxis, including ones being obtained by gene engineering methods; as well as in various educatory processes.

5 ex

 

The invention relates to biotechnology and can be used in genetic studies, in particular PCR diagnosis and genetic engineering construction, production of diagnostic test systems, in the creation of drugs for the prophylaxis of cholera, in the process of training especially dangerous infections.

Known avirulent test bacterial strain Vibrio cholerae of serovar 0139 (115 KM), not containing ctxAB genes used for immunological and genetic studies (RF patent N 2149898, IPC 12 N 1/00, 1/20 // (12 N 1/20, 12 R 1:63)).

The strain does not contain the ctxAB operon determining the production of cholera EN-crotoxin, causing hemolysis of sheep red blood cells and is avirulent for rabbit-suckers.

However, this strain does not belong to V to Vibrio cholerae 01 sarovara, biovars El tor circulation.

The closest to its taxonomic position is a strain of Vibrio cholerae biovars of eltor serovar Ogawa 156 used as a benchmark in the study of choleragenic (AU USSR N 1168598, IPC C 12 Q 1/20 C 12 R 1:63), which consistently causes choleragenic effect on the biomodels (rabbit-suckers) and promotes the accurate identification of Vibrio cholerae.

The distinguishing characteristics of a known strain of Vibrio cholerae biovars eitor of serovar Ogawa is a high virulence and toxicity to humans and rabbits-with whom the images, the ability to produce cholera enterotoxin. Information about the presence of pathogenicity genes are missing.

The objective of the invention is a test strain of Vibrio cholerae biovars of eltor serovar Ogawa, consistently keeping avirulent and toxigenic properties.

The inventive strain of Vibrio cholerae biovars eitor of serovar Ogawa isolated from a natural source - water open water on the territory of Turkmenistan.

The strain deposited in the Public collections of pathogenic bacteria at the Russian scientific research Institute "Microbe" (GKPB "M") under number 26 KM.

The strain of Vibrio cholerae biovars eitor of serovar Ogawa KM 26 is characterized by the following features.

Cultural morphological traits

Gram-negative, motile, slightly curved rods, with one polar located flagellum, spores and capsule form. On agar Martin and Hottinger (pH of 7.6 to 7.8) after 18 hours of growth at 37°With shape, round, translucent colonies with a bluish tint, with smooth edge, broth and 1% peptone water - uniform turbidity on the surface of a soft film.

Physiological and biochemical characteristics

Aerobe. Optimum temperature for growth is 37°C. the Optimum pH of 7.6 to 7.8. Produces endoperoxides belongs to 1 group of Heiberg, oxidizes and ferments glucose to acid without gas does not decompose the arabinose, dulcet, lactose, starch, decarbon iliret lysine, has no dihydrolase arginine, does not grow in the presence of 7-10% NaCl, analyzes sheep erythrocytes in a sample of Greig, agglutinated 2.5% Guinea pig erythrocytes, growing on medium with 30 units of polymyxin b, literoitca solid classical phage With, not Li-siruela phages El tor circulation and HDF - 3,4,5.

The strain of Vibrio cholerae the biovars eltor serovar Ogawa KM 26 agglutinated in the deployed agglutination reaction with holonymy diagnostic sera "On" and "Ogawa", not agglutinated holonymy diagnostic sera "Inaba" and "RO", not able to synthesize cholera enterotoxin and cause choleragen reaction when infected animals microorganism grown in various conditions, including the fermenter.

The special properties of the strain

The strain of Vibrio cholerae biovars eitor of serovar Ogawa KM 26 does not contain:

ctxA gene encoding the synthesis of the A-subunit of the cholera toxin (according to PCR with primers complementary to the nucleotide sequence of the ctxA gene and hybridization with ST-probe);

tcpA gene encoding the synthesis of the basic subunit toxicologically pilej adhesion (according to PCR with primers designed based on sequence tcpA gene);

the toxR gene responsible for the synthesis regulatory protein ToxR, coordinating the expression of ctx AB and t genes (according to PCR with primers to the toxR gene),

zot-the gene encoding the toxin synthesis zone is about absorption (according to PCR with primers on the sequence zot-rena).

The inventive strain of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 may be used:

1. In the production of PCR test systems for the detection of DNA of Vibrio cholerae (ctxA+) as a negative control for the test of finished products.

2. As avirulent test strain during differentiation of virulent strains of Vibrio cholerae biovars eltor by intestinal infection rabbit-suckers.

3. In immunological and genetic studies in the development of new living genetically engineered recombinant preparations for prophylaxis of cholera.

4. When demonstration course on preparation and improvement of specialists on especially dangerous infections on the basis of the anti-plague institutes, seminars for practitioners on the bases of anti-plague stations and departments of especially dangerous infections in epidemiological stations.

5. Avirulent test bacterial strain Vibrio cholerae biovars of eltor serovar Ogawa KM 26 can be used in quality control of manufactured V. agglutinating sera "Ogawa".

The use of strain is illustrated by the following examples.

Example 1, reflecting the use of strain as a negative control during production test systems, to confirm the specific activity and specificity of the produced test systems.

Production strain of Vibrio cholerae is Ivara classica 569B and strain for control of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 (158) are stored in the Public collections of pathogenic bacteria "Microbe" antiplague research Institute "Microbe". The control strain of V. cholerae KM 26 has a typical cultural-morphological and biochemical properties, is S-shaped, has no ctxA gene.

To obtain microbial suspensions of test strains vials of lyophilized culture of V. cholerae open sterile, and make 0.5 ml of 0.14 M NaCl solution (pH of 7.2). After dissolution in 1-3 min microbial suspension in a volume of 0.1 ml are seeded in 1% peptone water (pH of 7.6), incubated for 3-6 h at 37 ° °C. Then produce seed with biological loop # 2 on the agar of Hottinger pH of 7.6, incubated at 37°C for 18-20 hours

Obtaining DNA from strain. A suspension of microbial cells of Vibrio cholerae biovars classica 569B and Vibrio cholerae biovars of eltor serovar Ogawa KM 26, grown on agar of Hottinger, prepare 2 ml of 0.14 M NaCl (pH 7,2) industry standard sample turbidity 5 units gisk named after. Laurasia (CCA 42-28-P), which corresponds to 1.1×109M.K./ml of Vibrio cholerae. Then make a 10-fold dilution of strain V. cholerae V in 5 ml of 0.14 M solution of sodium chloride (pH of 7.2) to a concentration of 1×103, 1×102, 1×10 M.K/ml and Vibrio cholerae biovars of eltor serovar Ogawa 26 KM to 107M.K./ml

For disinfection to 5 ml of the bacterial suspensions of test strains add 0.5 ml of 0.1% solution of sodium merthiolate to a final concentration of 0.01%, is heated at a temperature of 56°in ECENA 40 minutes Then 1 ml of bacterial suspension from all dilutions separate pipettes and transferred into microtubes 1.5 ml and centrifuged at 8000 g for 15 min at a temperature of 20-25°C. the Precipitate resuspended in 50 ál of sterile deionized water, heated at a temperature of 100°C for 10 min, cooled for 2-3 min and centrifuged at 8000 g for 1 min at a temperature of 20-25°C. the Supernatant in an amount of 10 μl is used for PCR. Reaction amplification carried out according to the instructions attached to the test system. The results take into account the color of the DNA in the gel when viewed under UV light with a wavelength of 310 nm. If the sample DNA matrices Vibrio cholerae the biovars eltor serovar Ogawa KM 26 specific bands must be absent, and in the presence of DNA-matrices of Vibrio cholerae 569B should be visible fragments the size of 564 BP "Test-system for detection of DNA of Vibrio cholerae (ctxA+) by polymerase chain reaction" should be identified in the PCR 1000, M.K./ml Vibrio cholerae with ctxA gene and should not react with strains that do not contain ctxA gene.

The use of test strain of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 confirmed by specific activity and specificity of the test system. In a similar way the test strain of V. cholerae KM 26 can be used in the production of a multiplex PCR test C themes (tcp, tox R, zot genes).

Example 2. The use of the test strain during differentiation of virulent strains by intestinal infection biomodule.

The strain of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 when intestinal parasite infection rabbit-suckers in doses of 1×105, 1×107M.K./ml does not cause their death and at autopsy bioprobe animals no changes specific to choleragenic of the symptom, which indicates the inability of the strain to produce cholera toxin. Histological study of the organs and the lining of the intestine there is no marked submucosal edema shell, moderate activity APUD-system without quantitative changes. Quantitative indicators of goblet cells, bezepitelialnye lymphocytes integumentary epithelium did not differ from intact animals. Infection of the small rabbit-suckers virulent strain of Vibrio cholerae (V) caused their death when intestinal parasite infection in a dose of 1×105M.K./ml with a pronounced symptom, vidimie macroscopic changes of the internal organs, which confirms the ability of this strain to produce cholera enterotoxin.

Example 3. The use of strain in immunological and genetic research.

Avirulent test bacterial strain Vibrio cholerae biovars eitor of serovar Ogawa is M 26, not containing genes ctxA, tcpA, toxR, zot can be used as the basis for the design of recombinant live preparations for prophylaxis of cholera as appropriate "Criteria for assessment of vaccine strains V. cholerae" (guidelines, approved by the RF state Committee of sanitary, 1995, section 1.).

Example 4. The use of strain during the demonstration of the course on preparation and improvement of specialists on especially dangerous infections.

Avirulent strain of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 agglutinated holonymy sera "On" and "Ogawa", stably maintained S-shape, has a typical biochemical properties, which is very important for the training of specialists.

The use of avirulent test strain of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 can significantly improve the quality of training on especially dangerous infections, because you can avoid unexpected situations that may occur when working with virulent strains of V. cholerae. Similar strain for educational purposes at the present time there is no.

Example 5. The use of strain as a control output control sera "Ogawa".

The stability of the antigenic structure allows the use of strain as a control in the production of sera hall the situations agglutinating "Ogawa". Long-term storage of strains of Vibrio cholerae biovars of eltor serovar Ogawa KM 26 in dried condition at plus 4°does not affect antigenic stability.

When checking 80 strains of V. cholerae serovar Ogawa through the production of diagnostic agglutinating sera "Ogawa" 34 culture (42,5%) were agglutinability only this serum, and 46 strains (57,5%) agglutinative were serum "Ogawa" to titer, serum "Inaba" in dilutions of 1:10-1:50. The test strain of Vibrio cholerae biovars eitor of serovar Ogawa 26 KM in three replications with different batches of serum for 3 years (observation period) was agglutinable only serum "Ogawa", which indicates a stable antigenic structure.

Thus, the claimed strain has avirulent, toxygene properties, does not contain ctxA, tcpA, toxR, zot genes. The properties of the strain is stable and does not change during long-term storage, so you can use it in production PCT test systems, including multiplex PCT test systems, genetic, immunological studies. The examples clearly reflect the features of the claimed strain. The use of strain in the genetic, immunological studies can improve the quality of medical immunobiological preparations.

Avirulent test bacterial strain Vibrio cholerae biovars eltor Serov is RA Ogawa (ctxA -, tcpA-, toxR-, zot-)deposited in GKB "M" number 26 KM used in immunological and genetic research.



 

Same patents:

FIELD: biotechnology, medicine, veterinary, in particular drug for treatment and prevention of cow endometritis.

SUBSTANCE: claimed biological preparation contains dry biomass of strain Bifidobacterium pseudolongum N 533 and strain Lactobacillus salivarius N 117. Strains are cultivated separately on broth providing accumulation of necessary biomass. Biomass is concentrated by centrifugation and mixed with safety medium in ratio of 1:1-1:2, respectively. Obtained suspensions of B. pseudolongum N 533 and L. salivarius N 117 biomasses are blended in equal amounts, bottled into 2-4 ml bijous and dried.

EFFECT: preparation with high adhesion, antagonistic activity to opportunistic and pathogenic microflora and increased therapeutical effectiveness.

1 tbl, 2 ex

FIELD: biotechnology, medicine, veterinary, in particular drug for treatment and prevention of cow endometritis.

SUBSTANCE: claimed biological preparation contains dry biomass of strain Bifidobacterium pseudolongum N 533 and strain Lactobacillus salivarius N 117. Strains are cultivated separately on broth providing accumulation of necessary biomass. Biomass is concentrated by centrifugation and mixed with safety medium in ratio of 1:1-1:2, respectively. Obtained suspensions of B. pseudolongum N 533 and L. salivarius N 117 biomasses are blended in equal amounts, bottled into 2-4 ml bijous and dried.

EFFECT: preparation with high adhesion, antagonistic activity to opportunistic and pathogenic microflora and increased therapeutical effectiveness.

1 tbl, 2 ex

FIELD: biotechnology, microbiology, broth for bifidus bacteria cultivation.

SUBSTANCE: claimed broth for bifidus bacteria cultivation contains as main component pancreatic protein hydrolyzates of casein or forcemeat containing 700-900 mg% of amine nitrogen. Hydrolyzable mixture additionally contains 20-40 g/l of cow or pork liver waste. Broth also contains sodium chloride 4.0-5.0 g; L-cystine hydrochloride 0.10-0.15; microbiological agar 0.75-1.00 g; yeast autolyzate 300.0-400.0 ml; pancreatic casein hydrolyzate containing 700-900 mg% of amine nitrogen, diluted with distilled water up to amine nitrogen content of 140-160 mg%, 300.0-350.0 ml; pancreatic forcemeat hydrolyzate containing 700-900 mg% of amine nitrogen, diluted with distilled water up to amine nitrogen content of 140-160 mg%, 300.0-350.0 ml; 40 % lactose solution 25.0 ml.

EFFECT: new nutrient medium completely satisfying to bacterium population requirements.

2 cl, 3 tbl, 3 ex

FIELD: biotechnology and medical biology.

SUBSTANCE: invention relates to treatment preparation based on lactic bacteria of strain Lactobacillus acidophilus. Preparation contains 1.0-99.0 mass % of biomass of living bacteria Lactobacillus acidophilus N.V. Ep 317/402-X and balance: biomass of bacteria Lactobacillus acidophilus N.V. Ep 317/402-X, exposed to multiple freezing and thawing or autolysis, as source of cell detritus, extracellular and intracellular metabolites. Additionally preparation comprises 0.5-30.0 mg/ml of lysozyme and/or 0.001-0.1 mg/ml of nisin.

EFFECT: preparation with wide range of antagonistic activity, particularly in relation to pathogenic bacteria Helicobacter pylori associated with various forms of gastritis and duodenal and stomach ulcer, and high content of valuable substances.

2 cl, 2 tbl, 9 ex

FIELD: biotechnology.

SUBSTANCE: invention relates to nematophage fungi strain Duddingtonia flagrans useful as base in biological preparations producing. Strain Duddingtonia flagrans F-882 has activity against plant gall nematode and animal parasitic nematode, and stimulates plant growth and development.

EFFECT: strain with high plant growth and development stimulating activity and nematophage effect.

6 tbl, 2 dwg, 7 ex

FIELD: biotechnology and agriculture, in particular probiotic-containing compound feed for domestic animals, birds and fish.

SUBSTANCE: claimed additive is prepared by mixing of subtilis B-2250 and/or Bacillus licheniformis B-2252 biomass and carrier-sorbent and moisture capacitive filler as auxiliary substances. As carrier-sorbent hydrophilic and hydrophobic substances are used, and as moisture capacitive filler cation exchanging resins are used in predetermined ratio of dry mass. Probiotic additive in dry microcapsulated form is obtained by capillary sorptive drying of component mixture up to finished product humidity of 8-25 %.

EFFECT: probiotic additive with stable properties capable to improve feed conversion, to increase weight increment and to reduce animal mortality.

2 cl, 2 tbl, 4 ex

FIELD: microbiology.

SUBSTANCE: method for enzymatic conversion of phytosterol compositions to androstenedion (androst-4-ene-3,17-lion, AD) and androstadienedion (androsta-1,4-diene-3,17-dion, ADD) includes heating of phytosterol composition in presence of one or more solubilizers up to 100-130°C to form paste-like solution. Solubilizers are selected from polypropylene glycol, silicone or vegetable oil. Prepared paste-like mass is charged into bioreactor containing microorganism Mycobacterium MB 3683 and inorganic salt medium followed byfermentation. Method of present invention makes it possible to sufficiently increase of phytosterol composition concentration (30 g or more for 1 l of broth) and to increase produced product yield up to 80-90 %.

EFFECT: method of improved efficiency and yield.

6 cl, 1 dwg, 2 tbl, 5 ex

FIELD: biotechnology, in particular production of antibiotic pravastatin from compactin using microbiological transformation.

SUBSTANCE: microbiological compactin conversion to pravastatin is carried out by cultivation of strain Mortierella maculata n. sp. E-97 capable to hydrolyze pravastatin under deep anaerobic conditions. Broth for strain cultivation contains digestible carbon and nitrogen sources and mineral salts. Pravastatin is isolated from broth and purified. In claim of invention structural formulae of compactin (formula I) and pravastatin (formula II) are provided.

EFFECT: industrial scale method for pravastatin production.

56 cl, 1 dwg, 8 ex

FIELD: virology, veterinary.

SUBSTANCE: invention relates to thermosensitive strain of horse abortion herpes virus ENV-1, pharmaceutical composition, vaccine and method for animal immunization based on the same. Strain replication is limited by upper respiratory tract and doesn't cause of viremia. Furthermore strain has insufficient growth at temperature of wild-type virus normal growth. Strain of present invention, as well as compositions and vaccines based on the same are useful in clinical and viral horse protection against ENV-1 infections, viral induced abortion and paresis.

EFFECT: new pharmaceutical composition, vaccine and method for horse immunization.

4 cl, 10 tbl, 7 ex

FIELD: microbiology, medicine.

SUBSTANCE: invention relates to method of diagnosis and treatment of intestinal yersinioses and pseudotuberculosis. Claimed method includes Yersinia enterocolitica and Yersinia pseudotuberculosis isolation from clinic and other material. Broth (I-agar) contains (g/l): peptone 19.0-21.0; yeast extract 1.9-2.1; mannitol 19.0-21.0; sodium pyruvate 1.9-2.1; sodium chloride 0.95-1.05; magnesium sulfate 0.009-0.011; sodium deoxycholeta 0.48-0.53; neutral red 0.028-0.032; crystal violet 0.009-0.0011; agar-agar 11.9-13.1; irgasane 0.0039-0.0041; and balance: distilled water.

EFFECT: simplified broth composition with improved selectivity to Y. pseudotuberculosis without essential alteration of other medium characteristics.

3 tbl, 1 ex

FIELD: biotechnology, in particular pharmaceutical industry and medicine.

SUBSTANCE: method and kit for nuclear acid synthesis are disclosed. Said kit includes DNA-polymerase, catalyzing synthesis of complementary chain according to replacement mechanism, substrate therefore, two inner primer, and two oligonucleotide. Claimed method includes mixing of said kit with nuclear acid sample and mixture incubation at temperature sufficient for saving enzymatic activity of DNA-polymerase and stable coupling of sequence bases of inner primers with sequences being complementary thereto. Method of present invention makes it possible to obtain nuclear acid with nucleotide sequence wherein complementary nucleotide sequences are alternately bonded in one chain.

EFFECT: inexpensive method with increased synthesis specificity.

9 cl, 18 dwg, 8 ex

FIELD: genetic engineering, medicine.

SUBSTANCE: invention relates to T-cell receptor sequence being detected in patients with extended sclerosis and is useful in diagnosis and therapy. Oligonucleotide including sequence which represents or is derived from 5'-CTAGGGCGGGCGGGACTCACCTAC-3' or nucleotide sequence being fully complementary thereto. Oligonucleotide together with nuclear acid including nearly 15-30 oligonucleotides, which doesn't comprise oligonucleotide sequence and presents in region from Vβ to Jβ of Vβ13.1 gene in T-cell Vβ13.1-subgroup, wherein oligonucleotide and nuclear acid sequences don't present in the same chain of pair sequences of Vβ13.1 gene, is used in Vβ13.1 gene part amplification. In method for detection of LGRAGLTY motive, which is present in T-cell receptors of T-cell Vβ13.1-subgroup, oligonucleotide is used in combination with labeling particle. Once LGRAGLTY motive is detected, development monitoring and treatment are carried out by removing of LGRAGLTY motive-containing peptide.

EFFECT: simplified methods for detection of LGRAGLTY motive in T-cell receptors and treatment of patients with extended sclerosis.

21 cl, 7 dwg, 3 tbl, 3 ex

FIELD: medicine.

SUBSTANCE: method involves taking blood sample and dividing it plasma and blood cells fraction. The blood cells fraction is divided into erythrocytes and leukocytes and extracellular nucleic acids bound to erythrocytes and leukocytes surface are eluted. Extracellular nucleic acids fraction is separated and amplification analysis of not less than two known specific nucleic acid sequences associated to particular disease by applying appropriate method (polymerase chain reaction, ligase chain reaction, multiplex polymerase chain reaction and others).

EFFECT: high accuracy of diagnosis.

6 cl, 1 dwg, 4 tbl

FIELD: genetic engineering and medicine genetic.

SUBSTANCE: method for detection of gene resistance of subjects to infection by HIV1 is disclosed. Method includes DNA isolation, CCR5 gene PCR-amplification by using two primers complementary to two gene CCR5 sites. Further amplification products are restricted with endonuclease HincII (HindII), sizes of formed amplified fragments are determined, and on the base of obtained data deletion and single-nucleotide mutations are detected. Method of present invention makes it possible to simultaneously diagnose the presence of two mutation in human CCR5 gene, and (if individual has both mutation in heterozygote state) to distinguish cys- and trans-configurations.

EFFECT: method for diagnosis of congenital gene resistance to infection by HIV1.

2 dwg, 1 tbl, 2 ex

FIELD: molecular genetics and biotechnology.

SUBSTANCE: invention relates to method for detection of mismatched bases (point mutations) in nucleic acids. More particularly, strategy of phage selection bearing peptide analogs of functionally active protein modules of reparation system capable to selective binding to mismatched base sites on double-stranded DNA in certain locus is disclosed. Said strategy is based on combination of phage display by using biotinylated primers to provide PCR-fragments and paramagnetic particles with streptavidine to physically separate DNA-phage complex. Selected phages are selectively (preferably) bound to mismatched base in any heteroduplex composition. Present invention provides automatic method for molecular and epidemiological investigations of various diseases.

EFFECT: automatic method for molecular and epidemiological investigations.

4 cl, 2 dwg, 2 tbl, 4 ex

FIELD: medicine.

SUBSTANCE: method involves exposing genotype homozygous on functionally incomplete allele t receptor gene 1,25 dihydroxy vitamin D3 (VDR) with polymerase chain reaction being used with following sequencing electrophoresis in native polyacrylamide gel to predict predisposition. Odds ratio coefficient is calculated to predict disease clinical course severity. The coefficient shows how many times is severe degenerative dystrophic knee joint lesion occurrence probability higher in patients distinguished by unfavorable clinical course.

EFFECT: prevented pathological process aggravation.

2 cl

FIELD: food industry.

SUBSTANCE: for assay of soft wheat flour impurity in grit (semolina) of durum wheat polymerase chain reaction (PCR) is carried out for the presence of DNA sequence specific for genome of soft wheat (T. destivum). PCR is carried out using pair of oligonucleotide primers with homology to region of external transcribing spacer of highly repeating genes encoding ribosomal RNA in locus NorD3 belonging to genome D of soft wheat only. In the case the presence of corresponding DNA sequence of size 791 nulceotide pairs in products of PCR reaction the conclusion can be made about the presence of impurity ("adulteration") of grit with durum wheat flour (T. durum) with soft wheat. Invention can be used for assay of quality of macaroni articles and raw for their preparing.

EFFECT: improved assay method.

3 dwg

FIELD: molecular biology, medicine, pharmaceutical industry.

SUBSTANCE: invention discloses a method for identifying of definite functions of transcripts and proteins encoding by nucleic acid containing in the sample, a set for its realization comprising specific plasmid vector. Indicated vector comprises ori region, gene encoding tRNA and a cloning site located between two strong promoters optionally flanked by terminators and wherein the additional selective gene can be incorporated. Nucleic acid fragments obtained from a sample containing nucleic acid are fused into vector and vector is treated in vitro to obtain transcripts. Indicated transcripts are used in processes for testing their functions or for preparing proteins encoding by them. Proteins functions are tested also. A set for carrying out the method comprises, respectively, cellular extract for translation, agent for polymerization, agents that are necessary for testing one or some functions of proteins or transcripts encoding their, buffers, agents that are necessary for preparing nucleic acid fragments and specific vector. Applying invention provides rapid and effective carrying out function associated with polynucleotide sequence comprising in biological sample that involves nucleic acids.

EFFECT: improved identifying method.

27 cl, 7 dwg, 11 tbl

FIELD: medicine, hematology.

SUBSTANCE: one should isolate DNA out of peripheral blood lymphocytes due to polymerase chain reaction (PCR) technique of DNA synthesis, carry out genotyping for polymorphism of promoter area of TNF-alpha and TNF-beta gene. While detecting genotype LT*22 being characterized by availability of gene TNF-beta mutation in homozygous state, detect persons predisposed to the development of chronic lympholeukosis, and at certain combinations of genotypes TNF*22/LT*22 or TNF*12/LT*11 in patients with chronic lympholeukosis on should predict an aggressive flow of this disease.

EFFECT: higher accuracy of prediction.

3 ex, 3 tbl

FIELD: genetic engineering, biotechnology, biochemistry, agriculture, food industry, medicine.

SUBSTANCE: invention relates to the transformation of plant with nucleic acid encoding enzyme Δ6-desaturase in C. elegans that results to preparing a plant with enhanced content of gamma-linolenic acid and resistance to cold. Desaturase extracted from the plant can be used for preparing a drug used for treatment of disorder in body associated with deficiency of gamma-linolenic acid in it.

EFFECT: valuable biological properties of genes and desaturases.

36 cl, 9 dwg, 2 ex

FIELD: biotechnology, medicine, veterinary, in particular drug for treatment and prevention of cow endometritis.

SUBSTANCE: claimed biological preparation contains dry biomass of strain Bifidobacterium pseudolongum N 533 and strain Lactobacillus salivarius N 117. Strains are cultivated separately on broth providing accumulation of necessary biomass. Biomass is concentrated by centrifugation and mixed with safety medium in ratio of 1:1-1:2, respectively. Obtained suspensions of B. pseudolongum N 533 and L. salivarius N 117 biomasses are blended in equal amounts, bottled into 2-4 ml bijous and dried.

EFFECT: preparation with high adhesion, antagonistic activity to opportunistic and pathogenic microflora and increased therapeutical effectiveness.

1 tbl, 2 ex

Up!