Optimized recombinant protein encoding gene - human interferon beta analog

FIELD: biotechnology.

SUBSTANCE: nucleotide sequence encoding a recombinant protein - interferon beta analog, optimized for expression in E. coli cells. E. coli BL-30-B cell, intended for interferon beta recombinant protein production is obtained by transformation of E. coli BL (DE3) cells by rET30a plasmid containing the said nucleic acid. E. coli BL-32-B cell, intended for interferon beta recombinant protein production is obtained by transformation of E. coli BL (DE3) cells by rET32a plasmid containing the said nucleic acid.

EFFECT: invention provides a recombinant protein.

3 cl, 5 dwg, 9 tbl, 9 ex

 



 

Same patents:

FIELD: biotechnology.

SUBSTANCE: strain of bacteria Rhodococcus sp. LER-12 having the ability to dispose quickly of crude oil and petroleum products (diesel fuel, motor oil, hydraulic oil, gas condensate) is deposited in the RNCM under the registration number Rhodococcus sp. RNCM Ac-2626D and can be used for purification of soils and water from contaminations with crude oil and petroleum products, in a wide temperature range from +4 to +30°C.

EFFECT: invention enables to improve the quality of purification of soil and water from crude oil and petroleum products.

3 tbl, 5 ex

FIELD: biotechnology.

SUBSTANCE: strain is deposited in the State collection of pathogenic microorganisms "SCIM-Obolensky" under the number F-11. The culture fluid centrate obtained as a result of culturing of said the strain on the liquid nutrient medium has high activity against anthrax causative agent. The zone of inhibition of growth of Bacillus anthracis is 20-25 mm and 35-45 mm when the culture fluid centrate of the said strain is used in an amount of 10 mcl and 100 mcl, respectively.

EFFECT: strain of micromycete Trichoderma hamatum has antibacterial activity against anthrax causative agent Bacillus anthracis.

4 tbl, 3 ex

FIELD: biotechnology.

SUBSTANCE: nutrient medium comprises sodium chloride, potassium chloride, magnesium sulphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, calcium chloride, sodium bicarbonate, L-arginine, L-glutamine, L-tyrosine, L-tryptophan, calcium pantothenate, pyridoxal HCl, thiamine, meso-inositol, nicotinamide, riboflavin, folic acid, glucose, enzymatic hydrolyzate of muscle proteins, lactalbumin enzymatic hydrolyzate, sodium salt of benzylpenicillin with activity of 0.09×105-1.1×105, kanamycin sulphate with activity of 0.09×105-1.1×105, nystatin with activity of 6×104 to 6.5×104 IU/l, choline chloride, succinic acid and distilled water in a predetermined ratio of components.

EFFECT: invention enables to improve the quality of the product obtained.

1 tbl, 1 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology. The strain of fungus Aspergillus niger B-6 is the producer of citric acid. The strain Aspergillus niger is deposited in the joint Russian Collection of Agrocultural Microorganisms RAAS (RCAM) under registration number RCAM02149 and can be applied in the production of citric acid.

EFFECT: invention makes it possible to increase the output of citric acid.

2 tbl, 6 ex

FIELD: biotechnologies.

SUBSTANCE: method of production of nanodisperse additive for concrete is offered. Cyanobacteria of the sort Pseudanabaena sp. 0411 or Leptolyngbya laminosa 0412 are cultivated in a nutrient medium at the temperature 23-25°C. The nutrient medium is Z-8 medium with adding of sodium silicate solution neutralized by 2 M HCl. Ratio of solution of silicate of sodium and Z-8 5:1 medium. Cultivation is carried out in the bioreactor at continuous lighting and mixing during 10 days with the subsequent removal of nutrient medium residue. Then the culture is poured by 30% hydrogen peroxide solution and heated up to 70°C. The produced biosilicated nanotubes are washed out by distilled water with subsequent treatment in a mechanical activator in the water medium of anion surface-active substance of naphthalene formaldehyde type at the ultrasound frequency 35 kHz and concentration of solid phase 5% up to the particle size 85-250 nanometres.

EFFECT: increase of mobility of concrete mix, curing acceleration, increase of strength and density of concrete, decrease of water absorption.

3 tbl, 1 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanovermiculite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanobentonite and distilled water.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 14 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nano sapropel and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of phosphate-mobilising and nitrogen-fixing microorganisms.

1 tbl, 15 ex

FIELD: biotechnologies.

SUBSTANCE: nutritive medium contains potassium dihydrophosphate, potassium hydrophosphate, magnesium sulphate heptahydrate, sodium chloride, calcium sulphate dihydrate, sodium molybdate, iron (II) sulphate, saccharose, nanozeolite and distilled water with a defined component ratio.

EFFECT: enhanced growth rate of nitrogen-fixing and phosphate-mobilising microorganisms.

1 tbl, 15 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology and concerns the strain Escherichia coli BL21(DE3)Gold/pETmin-CypA producing human recombinant cyclophilin A. The characterised strain is produced by cell transformation of the strain BL21(DE3)Gold by pETmin-CypA plasmid. The plasmid has a size of 5865 base pairs and contains XhoI-NdeI fragment of pET-22b(+) vector carrying ampR ampicillin resistance gene, a promoter and RNA-polymerase T7 phage terminator and a polylinker. A fragment produced by PCR with using the oligonucleotide primers 5'-TTATACATATGGTCAACCCGACCGTGTTCTTC and 5'-TTTCTCGAGTTATTCGAGTTGTCCACAGTCAGC of pCyPAwt/pGEX-2TK plasmid with the closed fragment of hrCpA (human recombinant cyclophilin A) gene having a size of 498 base pairs is cloned at XhoI-NdeI sites.

EFFECT: presented strain is high-producing; it requires no complicated clean-up system and can be used in cancer treatment.

2 dwg

FIELD: chemistry.

SUBSTANCE: invention relates to the field of biotechnology and can be applied for the selection of affine molecules. Constructed is a plasmid DNA pGST/APT/X 6193 bp long, with the weight of 4.09 MDa, containing as a genetic marker the gene β-lactase and unique sites of the recognition of restriction endonucleases, located at the following distance to the right from the site NdeI: BgIII 665bp, BamHI 779 bp., XhoI 801 bp. The plasmid pGST/APT/X consists of a NdeI-XhoI DNA fragment of the commercial plasmid pET22b(+) and nucleotide sequence SEQ ID NO:1, inserted by sites NdeI-XhoI and combining in its composition the sequences, which code a fragment of a gene of glutathione-S-transferase, a peptide substrate of a lethal factor of anthrax RRKKVYPYPME, peptide GGLNDIFEAQKIEWHED, biotinylated in vivo under the influence of E.coli biotin-ligase, and a linker sequence, substituted by genetic-engineering manipulations by sites of restriction endonucleases BamHI and XhoI for a sequence, coding a target protein B-subunit of shiga-toxin I.

EFFECT: invention makes it possible to carry out the selection of highly specific aptamers to the said target protein.

2 cl, 4 dwg, 4 ex

FIELD: chemistry.

SUBSTANCE: invention relates to biotechnology, the modified polypeptide having homoserine-O-acetyltransferase activity having the amino acid sequence of SEQ ID NO:17 or at least 95% homologous thereto, in which the amino acid at position 111 from the start amino acid methionine, of the sequence is substituted with glutamic acid. The invention also relates to a method for producing O-acetyl homoserine, comprising culturing the microorganism belonging to the genus Escherichia, transformed with polynucleotide, encoding the said polypeptide.

EFFECT: high production yield of O-acetyl homoserine.

18 cl, 5 dwg, 3 tbl, 5 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: inventions concerns recombinant DNA pA3, recombinant plasmid DNA pQE 30-pA3, strain E.coli M 15-A3, recombinant polypeptide A3, test systems for the qualitative detection and quantitative determination of microalbuminuria. The characterised recombinant DNA pA3 is produced by a polymerase chain reaction with the use of chromosomal DNA of the strain DG 13 of group B streptococci and structured primers. An amplification fragment has been cloned by a system of expression vectors pQE-pQE 30 in E.coli M 15 to produce recombinant DNA pQE 30-pA3, which codes an amino acid sequence of the recombinant polypeptide A3 having an ability to bind human serum albumin (HSA) selectively.

EFFECT: presented inventions can be used in the medical practice and industry for producing the test systems for the detection and quantitative determination of microalbuminuria - one of the earliest signs of renal irritation in the patients suffering from diabetes mellitus and essential arterial hypertension.

6 cl, 9 dwg, 1 tbl, 8 ex

FIELD: medicine.

SUBSTANCE: invention refers to biotechnology and can be used for recombinant production of human tissue factor (hTF). Constructed is a plasmid pHYP-10ETFCS6 having a length of 5,912 b.p. with a physical map presented on Fig. 2, for expression in a bacterium of the genus Escherichia, which is a precursor of the mutant [C245S] hTF containing an inseparable N-terminal leader peptide containing a deca-histidine cluster and an enterokinase identification sequence fused in a frame with a sequence coding the above mutein fused in the frame with the sequence coding the additional inseparable C-terminal peptide containing the deca-histidine cluster. A method for producing the precursor of the mutein[C245S]hTF contains culturing the producing bacterium in a nutrient medium, recovering inclusion bodies, solubilising the precursor protein, performing a metal chelator chromatography in the denaturation environment, re-folding and diafiltration of the protein solution. A method for producing the mature mutein[C245S] hTF involves detecting the N-terminal leader peptide from the above mutein precursor with using enterokinase and recovering the target protein.

EFFECT: invention enables increasing the level of biosynthesis and yield of pro-coagulation active hTF.

9 cl, 5 dwg, 1 tbl, 7 ex

FIELD: chemistry.

SUBSTANCE: claimed inventions deal with an isolated polynucleotide, coding a polypeptide, involved in biosynthesis of pyripyropene A, a vector and a host cell, including such a polypeptide, and methods of obtaining pyripyropene A precursors, including the host cell cultivation. The claimed polynucleotide codes the polypeptide, possessing any one or more of the activities - polyketide synthase, prenyltransferase, hydroxylase, acetyltransferase or adenylate synthase.

EFFECT: claimed inventions make it possible to synthesise pyripyropene A, which is an insecticidal agent, and can be used in the formation of plant resistance to pest insects.

16 cl, 11 dwg, 1 tbl, 11 ex

FIELD: chemistry.

SUBSTANCE: invention relates to field of biotechnology, in particular to obtaining recombinant enzyme-labelled antigen G2 of hantavirus Dobrava. Essence of invention consists in the following: claimed method of obtaining antigen G2 of hantavirus Dobrava consists in expression of antigen in cells of E.coli in form of enzyme-labelled antigen of G2 hantaviruses based on HT protein antigen. Beta-galactosidase, which is highly active stable enzyme, serves enzyme label for protein antigen. Invention can be used to increase specificity and reproducibility of immunosorbent assay in HFRS diagnostics.

EFFECT: presence of commercially available chromogenic substrate of beta-galactosidase (X-gal) makes it possible to quickly estimate result of immunosorbent reaction visually or by change of optic density of solution in visible area.

6 dwg

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biochemistry and represents a new bifunctional PSH protein containing human peroxiredoxin Prx6 and manganese superoxide dismutase MnSOD possessing the antioxidant activity of superoxide dismutase and peroxidase. What is also described is a chimeric nucleic acid coding the presented protein. A method for preparing the presented protein by culturing cells of the strain E.coli BL21(PSH) transformed by constructed recombinant expression vector based on pET22b(+) plasmid is disclosed.

EFFECT: invention enables producing high-yield protein PSH possessing the high antioxidant activity.

4 cl, 6 dwg, 3 tbl, 7 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention relates to field of biotechnology, namely to recombinant obtaining G-CSF, and can be used for production of G-CSF in cells of E.coli. For effective production of protein in cells of E.coli G-CSF-coding DNA sequence is optimised. On the basis of obtained optimised DNA sequence plasmid pAS017, also including NdeI/BamHI-fragment of DNA of pETM-50 vector and having physical map, presented on the drawing, is constructed.

EFFECT: invention provides effective production of protein in cells of Ecoli.

2 cl, 1 dwg, 1 tbl, 3 ex

FIELD: medicine, pharmaceutics.

SUBSTANCE: invention refers to biotechnology, particularly to genetically engineered production of human proteins, and may be used for preparing human epidermal growth factor (hEGF) in bacterial cells in the form of glutathione-3-transferase fusion protein. What is constructed is the recombinant DNA coding GST-hEGF fusion protein which consists of an amino acid sequence of glutathione-S-transferase and an amino acid sequence of human epidermal growth factor divided by a cleavage site by enterokinase, and characterised by the nucleotide sequence SEQ ID NO:1. The KpnI/XhoI fragment of the vector pET41 and the above recombinant DNA are used to create the recombinant plasmid pAS007 for expression of GST-hEGF fusion protein in E.coli cells.

EFFECT: invention enables reaching high GST-hEGF expression levels in Ecoli cells.

2 cl, 3 dwg, 1 tbl, 5 ex

FIELD: chemistry.

SUBSTANCE: group of inventions relates to biotechnology, gene and protein engineering and specifically to recombinant plasmid DNA pG1-Rm7, which facilitates synthesis of hybrid protein G1-Rm7 in Escherichia coli cells, which is capable of biding the tumour necrosis factor and has bioluminescence of luciferase Renilla muelleri, where said plasmid DNA includes the nucleotide sequence SEQ ID NO: 1 and can be in medicine. The invention also relates to the protein pG1-Rm7 having molecular weight of 65.4 kDa, consisting of a single-strand anti tumour necrosis factor antibody, a GGSGGS peptide and modified luciferase Renalla muelleri and characterised by SEQ ID NO: 2.

EFFECT: invention enables to obtain a highly sensitive reporter for detecting a tumour necrosis factor via bioluminescent analysis.

2 cl, 4 dwg, 3 ex

FIELD: chemistry.

SUBSTANCE: invention relates to compositions of active cytokine - interferone-gamma, and to cost-effective methods of obtaining the claimed compositions and of treatment and immune reaction acceleration. The methods involve introduction of alternated recombinant cell (ARC) represented by P. Fluorescens and including heterologic gene of interferone-gamma.

EFFECT: increased specific activity of interferone.

22 cl, 10 ex, 13 tbl, 13 dwg

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