[go: up one dir, main page]

RU2016123389A - Plasmid vector pET-mChBac75Na, strain of the bacterium Eschrichia coli BL (DE3 / pET-mChBac75Na for expression of the antimicrobial peptide of minibactenecin ChBac7Na and a method for producing said peptide - Google Patents

Plasmid vector pET-mChBac75Na, strain of the bacterium Eschrichia coli BL (DE3 / pET-mChBac75Na for expression of the antimicrobial peptide of minibactenecin ChBac7Na and a method for producing said peptide Download PDF

Info

Publication number
RU2016123389A
RU2016123389A RU2016123389A RU2016123389A RU2016123389A RU 2016123389 A RU2016123389 A RU 2016123389A RU 2016123389 A RU2016123389 A RU 2016123389A RU 2016123389 A RU2016123389 A RU 2016123389A RU 2016123389 A RU2016123389 A RU 2016123389A
Authority
RU
Russia
Prior art keywords
mchbac75na
pet
minibactenecin
his8
peptide
Prior art date
Application number
RU2016123389A
Other languages
Russian (ru)
Other versions
RU2618850C2 (en
Inventor
Сергей Владимирович Баландин
Илья Александрович Болосов
Павел Валерьевич Пантелеев
Владимир Николаевич Кокряков
Ольга Валерьевна Шамова
Татьяна Владимировна Овчинникова
Original Assignee
Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН)
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) filed Critical Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН)
Priority to RU2016123389A priority Critical patent/RU2618850C2/en
Publication of RU2016123389A publication Critical patent/RU2016123389A/en
Application granted granted Critical
Publication of RU2618850C2 publication Critical patent/RU2618850C2/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C12N15/72—Expression systems using regulatory sequences derived from the lac-operon
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2330/00—Production
    • C12N2330/50—Biochemical production, i.e. in a transformed host cell
    • C12N2330/51—Specially adapted vectors
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2510/00—Genetically modified cells
    • C12N2510/02—Cells for production
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2800/00—Nucleic acids vectors
    • C12N2800/22—Vectors comprising a coding region that has been codon optimised for expression in a respective host

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Claims (5)

1. Плазмидный вектор pET-mChBac75Na для экспрессии в клетках Escherichia coli антимикробного пептида минибактенецина ChBac7.5Nα в составе гибридного белка His8-TrxL-mChBac75Nα, состоящий из двух фрагментов ДНК:1. Plasmid vector pET-mChBac75Na for expression in Escherichia coli cells of the antimicrobial peptide minibactenecin ChBac7.5Nα in the fusion protein His8-TrxL-mChBac75Nα, consisting of two DNA fragments: BglII/XhoI-фрагмента с нуклеотидной последовательностью SEQ ID No. 2, содержащего промотор транскрипции Т7 РНК-полимеразы, lac-оператор, участок связывания рибосомы и участок, кодирующий октагистидиновую последовательность, белок тиоредоксин и минибактенецин ChBac7.5Nα;BglII / XhoI fragment with the nucleotide sequence of SEQ ID No. 2, comprising a T7 RNA polymerase transcription promoter, a lac operator, a ribosome binding site and a section encoding an octa histidine sequence, a thioredoxin protein and ChBac7.5Nα minibactenecin; BglII/XhoI-фрагмента плазмиды рЕТ-20b(+), содержащего терминатор транскрипции Т7 РНК-полимеразы, сайт инициации репликации и ген β-лактамазы.The BglII / XhoI fragment of plasmid pET-20b (+) containing the T7 RNA polymerase transcription terminator, a replication initiation site, and a β-lactamase gene. 2. Штамм бактерии Escherichia coli BL21(DE3)/pET-mChBac75Na - продуцент гибридного белка His8-TrxL-mChBac75Nα, получаемый путем трансформации клеток родительского штамма BL21(DE3) плазмидным вектором pET-mChBac75Na по п. 1.2. The bacterial strain Escherichia coli BL21 (DE3) / pET-mChBac75Na is a producer of the His8-TrxL-mChBac75Nα hybrid protein obtained by transforming cells of the parent strain BL21 (DE3) with the plasmid vector pET-mChBac75Na according to claim 1. 3. Способ получения антимикробного пептида минибактенецина ChBac7.5Nα, включающий экспрессию гена гибридного белка His8-TrxL-mChBac75Nα в штамме-продуценте Escherichia coli BL21(DE3)/pET-mChBac75Na по п. 2, лизис клеток, аффинную очистку гибридного белка His8-TrxL-mChBac75Nα на металлохелатном носителе в денатурирующих условиях, расщепление гибридного белка His8-TrxL-mChBac75Nα бромцианом в 6М гидрохлориде гуанидина и очистку целевого пептида методом обращенно-фазовой ВЭЖХ.3. A method of producing an antimicrobial peptide of minibactenecin ChBac7.5Nα, comprising expressing the His8-TrxL-mChBac75Nα fusion protein gene in the Escherichia coli BL21 (DE3) / pET-mChBac75Na producer strain of claim 2, cell lysis, His8 Trx fusion protein fusion lysis -mChBac75Nα on a metal chelate support under denaturing conditions, cleavage of the His8-TrxL-mChBac75Nα fusion protein with cyanogen bromide in 6M guanidine hydrochloride and purification of the target peptide by reverse-phase HPLC.
RU2016123389A 2016-06-14 2016-06-14 pET-mChBac75Na PLASMID VECTOR, ESCHRICHIA COLI BL21(DE3/ pET-mChBac75Na BACTERIA STRAINS FOR CHBAC7NA MINIBACTENECIN ANTIMICROBIAL PEPTIDE EXPRESSION AND METHODS FOR PRODUCTION OF THESE PEPTIDES RU2618850C2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
RU2016123389A RU2618850C2 (en) 2016-06-14 2016-06-14 pET-mChBac75Na PLASMID VECTOR, ESCHRICHIA COLI BL21(DE3/ pET-mChBac75Na BACTERIA STRAINS FOR CHBAC7NA MINIBACTENECIN ANTIMICROBIAL PEPTIDE EXPRESSION AND METHODS FOR PRODUCTION OF THESE PEPTIDES

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
RU2016123389A RU2618850C2 (en) 2016-06-14 2016-06-14 pET-mChBac75Na PLASMID VECTOR, ESCHRICHIA COLI BL21(DE3/ pET-mChBac75Na BACTERIA STRAINS FOR CHBAC7NA MINIBACTENECIN ANTIMICROBIAL PEPTIDE EXPRESSION AND METHODS FOR PRODUCTION OF THESE PEPTIDES

Publications (2)

Publication Number Publication Date
RU2016123389A true RU2016123389A (en) 2017-01-10
RU2618850C2 RU2618850C2 (en) 2017-05-11

Family

ID=57955802

Family Applications (1)

Application Number Title Priority Date Filing Date
RU2016123389A RU2618850C2 (en) 2016-06-14 2016-06-14 pET-mChBac75Na PLASMID VECTOR, ESCHRICHIA COLI BL21(DE3/ pET-mChBac75Na BACTERIA STRAINS FOR CHBAC7NA MINIBACTENECIN ANTIMICROBIAL PEPTIDE EXPRESSION AND METHODS FOR PRODUCTION OF THESE PEPTIDES

Country Status (1)

Country Link
RU (1) RU2618850C2 (en)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
RU2721273C1 (en) * 2019-04-22 2020-05-18 Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) Nikomycin peptide from nereid nicomache minor possessing antimicrobial and antitumour activity

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9044438B2 (en) * 2005-06-17 2015-06-02 Yitzchak Hillman Disease treatment via antimicrobial peptides or their inhibitors
RU2580031C2 (en) * 2015-03-16 2016-04-10 Федеральное государственное бюджетное учреждение науки институт биоорганической химии им. академиков М.М. Шемякина и Ю.А. Овчинникова Российской академии наук (ИБХ РАН) PLASMID VECTOR pET-His8-TrxL-Acip1 STRAIN OF BACTERIA Escherichia coli BL21(DE3)/pET-His8-TrxL-Acip1 FOR EXPRESSION OF ANTIMICROBIAL PEPTIDE ACIPENSINA-1 AND METHOD OF PRODUCING SAID PEPTIDE

Also Published As

Publication number Publication date
RU2618850C2 (en) 2017-05-11

Similar Documents

Publication Publication Date Title
US11261219B2 (en) Proteolytic inactivation of select proteins in bacterial extracts for improved expression
Lipinszki et al. Enhancing the translational capacity of E. coli by resolving the codon bias
Eschenfeldt et al. New LIC vectors for production of proteins from genes containing rare codons
Wu et al. Combining CRISPR and CRISPRi systems for metabolic engineering of E. coli and 1, 4-BDO biosynthesis
JP2017513479A5 (en)
US10696963B2 (en) Selective optimization of a ribosome binding site for protein production
WO2018017828A1 (en) Biocatalyst comprising photoautotrophic organisms producing recombinant enzyme for degradation of harmful algal bloom toxins
RU2016123389A (en) Plasmid vector pET-mChBac75Na, strain of the bacterium Eschrichia coli BL (DE3 / pET-mChBac75Na for expression of the antimicrobial peptide of minibactenecin ChBac7Na and a method for producing said peptide
CN115747187B (en) Recombinase UvsX and expression gene and application thereof
RU2015109081A (en) PLASMID VECTOR pET-His8-TrxL-AciP1, STRAIN OF THE BACTERIA Eschrichia coli BL21 (DE3) / pET-His8-TrxL-AciP1 FOR EXPRESSION OF ANTIMICROBIC ACEPEPSIN-1 PEPTIDE AND METHOD OF SEARCH
Kurata et al. Identification of the set of genes, including nonannotated morA, under the direct control of ModE in Escherichia coli
CN103667331B (en) Recombinase gene bet is as a kind of application of intestinal bacteria heterologous protein expression fusion tag
WO2012177655A3 (en) Methods and systems for tracking bioremediation processes
Zhang et al. Assay and characterization of an osmolarity inducible promoter newly isolated from Bacillus subtilis
Chhetri et al. Recombinant expression, purification and preliminary characterization of the mRNA export factor MEX67 of Saccharomyces cerevisiae
Xu et al. Cytoplasmic Overexpression, Folding, and Processing of Penicillin Acylase Precursor in Escherichiacoli
CN103710373B (en) A kind of prokaryotic fusion expression vector and application thereof
ES2897017B2 (en) RELAXASE-CAS CHIMERIC PROTEIN
RU2009126233A (en) Recombinant plasmid DNA pQe30_PS-CFP2 / TurboYFP_MBP7, coding fragment of human myelin basic protein fused 80-104 IN THE COMPOSITION OF STRUCTURES with yellow and cyan fluorescent protein EPeFRET, STRAIN Escherichia coli BL21 (DE3) - PRODUCER said protein, AND METHOD OF PRODUCING RECOMBINANT PROTEIN EPeFRET
Afful et al. Overproduction and purification of highly active recombinant Pseudomonas aeruginosa str. PAO1 RNA polymerase holoenzyme complex
RU2010127458A (en) Recombinant plasmid DNA pPA-OPRF, encodes SYNTHESIS OF RECOMBINANT PROTEIN F OUTER MEMBRANE PSEUDOMONAS AERUGINOSA, STRAIN ESCHERICHIA coli PA-OPRF-RECOMBINANT PROTEIN PRODUCER F OUTER MEMBRANE PSEUDOMONAS AERUGINOSA, AND METHOD OF PRODUCING RECOMBINANT PROTEIN F OUTER MEMBRANE PSEUDOMONAS AERUGINOS
RU2016129503A (en) Plasmid vector pET-pPsLTP, strain of the bacterium Escherichia coli BL21 (DE3) Star / pET-pPsLTP-producer of food allergen of peas Pis s 3 and method for producing said allergen
RU2015139726A (en) Recombinant plasmid DNA pER-APHC3 encoding a fusion protein capable of autocatalytic cleavage to form APHC3, Eschrichia coli strain C3030 / pER-APHC3 producing these proteins and a method for producing recombinant APCH3
CN106811476B (en) A set of vectors for the expression of multiple protein complexes and its application
RU2011142855A (en) Recombinant plasmid DNA pPA-OPRFI, encodes the synthesis of a fusion recombinant protein F-I outer membrane PSEUDOMONAS AERUGINOSA, STRAIN ESCHERICHIA COLI PA-OPRFI-PRODUCER HYBRID RECOMBINANT PROTEIN F-I outer membrane PSEUDOMONAS AERUGINOSA, AND METHOD FOR PRODUCING HYBRID RECOMBINANT PROTEIN F-I outer membrane PSEUDOMONAS AERUGINOSA