CN113876952A - 氮源用于抑制细菌三型分泌系统的应用 - Google Patents
氮源用于抑制细菌三型分泌系统的应用 Download PDFInfo
- Publication number
- CN113876952A CN113876952A CN202010631326.XA CN202010631326A CN113876952A CN 113876952 A CN113876952 A CN 113876952A CN 202010631326 A CN202010631326 A CN 202010631326A CN 113876952 A CN113876952 A CN 113876952A
- Authority
- CN
- China
- Prior art keywords
- nitrogen source
- pathogenic bacteria
- host
- bacteria
- pathogenic
- Prior art date
- Legal status (The legal status 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 status listed.)
- Granted
Links
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 title claims abstract description 76
- 229910052757 nitrogen Inorganic materials 0.000 title claims abstract description 38
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 12
- 108010069584 Type III Secretion Systems Proteins 0.000 title claims abstract description 8
- 230000001580 bacterial effect Effects 0.000 title abstract description 18
- 208000015181 infectious disease Diseases 0.000 claims abstract description 20
- 230000001717 pathogenic effect Effects 0.000 claims abstract description 16
- 239000003814 drug Substances 0.000 claims abstract description 15
- 244000052769 pathogen Species 0.000 claims abstract description 4
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 32
- 235000004554 glutamine Nutrition 0.000 claims description 32
- 241000588724 Escherichia coli Species 0.000 claims description 31
- 244000052616 bacterial pathogen Species 0.000 claims description 30
- 241000894006 Bacteria Species 0.000 claims description 25
- 108090000623 proteins and genes Proteins 0.000 claims description 17
- 238000000034 method Methods 0.000 claims description 15
- 230000014509 gene expression Effects 0.000 claims description 14
- 102000004169 proteins and genes Human genes 0.000 claims description 14
- 235000018102 proteins Nutrition 0.000 claims description 13
- 241000589615 Pseudomonas syringae Species 0.000 claims description 9
- 230000003248 secreting effect Effects 0.000 claims description 9
- 230000028327 secretion Effects 0.000 claims description 9
- 230000001018 virulence Effects 0.000 claims description 7
- 241000588923 Citrobacter Species 0.000 claims description 6
- 206010061217 Infestation Diseases 0.000 claims description 6
- 230000002008 hemorrhagic effect Effects 0.000 claims description 6
- 241000588722 Escherichia Species 0.000 claims description 5
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 5
- 241001465754 Metazoa Species 0.000 claims description 5
- DCXYFEDJOCDNAF-UHFFFAOYSA-N Asparagine Chemical compound OC(=O)C(N)CC(N)=O DCXYFEDJOCDNAF-UHFFFAOYSA-N 0.000 claims description 4
- 208000035143 Bacterial infection Diseases 0.000 claims description 4
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 claims description 4
- 241000282414 Homo sapiens Species 0.000 claims description 4
- 235000013922 glutamic acid Nutrition 0.000 claims description 4
- 239000004220 glutamic acid Substances 0.000 claims description 4
- 208000022362 bacterial infectious disease Diseases 0.000 claims description 3
- 230000000369 enteropathogenic effect Effects 0.000 claims description 3
- 230000029142 excretion Effects 0.000 claims description 3
- 235000013305 food Nutrition 0.000 claims description 3
- 239000003112 inhibitor Substances 0.000 claims description 3
- 125000001477 organic nitrogen group Chemical group 0.000 claims description 3
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 claims description 2
- 150000003863 ammonium salts Chemical group 0.000 claims description 2
- 235000009582 asparagine Nutrition 0.000 claims description 2
- 229960001230 asparagine Drugs 0.000 claims description 2
- 235000003704 aspartic acid Nutrition 0.000 claims description 2
- OQFSQFPPLPISGP-UHFFFAOYSA-N beta-carboxyaspartic acid Natural products OC(=O)C(N)C(C(O)=O)C(O)=O OQFSQFPPLPISGP-UHFFFAOYSA-N 0.000 claims description 2
- 125000000404 glutamine group Chemical group N[C@@H](CCC(N)=O)C(=O)* 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 150000002823 nitrates Chemical class 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims description 2
- 230000032258 transport Effects 0.000 claims description 2
- 239000012459 cleaning agent Substances 0.000 claims 2
- 239000000654 additive Substances 0.000 claims 1
- 230000000996 additive effect Effects 0.000 claims 1
- 239000003674 animal food additive Substances 0.000 claims 1
- 239000002778 food additive Substances 0.000 claims 1
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 description 35
- 241000699670 Mus sp. Species 0.000 description 21
- 239000003242 anti bacterial agent Substances 0.000 description 17
- 229940088710 antibiotic agent Drugs 0.000 description 17
- 239000001963 growth medium Substances 0.000 description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 12
- 230000003115 biocidal effect Effects 0.000 description 10
- 239000007788 liquid Substances 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 239000008223 sterile water Substances 0.000 description 9
- 238000012258 culturing Methods 0.000 description 8
- 230000002829 reductive effect Effects 0.000 description 8
- 229940079593 drug Drugs 0.000 description 6
- 230000012010 growth Effects 0.000 description 6
- 239000002609 medium Substances 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 206010059866 Drug resistance Diseases 0.000 description 5
- 229930182816 L-glutamine Natural products 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 235000005911 diet Nutrition 0.000 description 5
- 239000003651 drinking water Substances 0.000 description 5
- 235000020188 drinking water Nutrition 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 3
- 241000219195 Arabidopsis thaliana Species 0.000 description 3
- 238000009631 Broth culture Methods 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 239000006137 Luria-Bertani broth Substances 0.000 description 3
- 241000589626 Pseudomonas syringae pv. tomato Species 0.000 description 3
- 235000001014 amino acid Nutrition 0.000 description 3
- 229940024606 amino acid Drugs 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 239000012228 culture supernatant Substances 0.000 description 3
- 230000037213 diet Effects 0.000 description 3
- 210000003608 fece Anatomy 0.000 description 3
- 231100000518 lethal Toxicity 0.000 description 3
- 230000001665 lethal effect Effects 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- 239000006228 supernatant Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108020004513 Bacterial RNA Proteins 0.000 description 2
- 108010078791 Carrier Proteins Proteins 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000589516 Pseudomonas Species 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000000378 dietary effect Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- GNBHRKFJIUUOQI-UHFFFAOYSA-N fluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 GNBHRKFJIUUOQI-UHFFFAOYSA-N 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 210000000936 intestine Anatomy 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 230000026533 negative regulation of protein secretion Effects 0.000 description 2
- -1 nitrogen-containing compound Chemical class 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000012827 research and development Methods 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 241001453380 Burkholderia Species 0.000 description 1
- 101100298998 Caenorhabditis elegans pbs-3 gene Proteins 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 241000606161 Chlamydia Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 241000588921 Enterobacteriaceae Species 0.000 description 1
- 241001646719 Escherichia coli O157:H7 Species 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000588748 Klebsiella Species 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 101100451485 Pseudomonas syringae pv. syringae hrpL gene Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 108700008625 Reporter Genes Proteins 0.000 description 1
- 241000607142 Salmonella Species 0.000 description 1
- 241000607768 Shigella Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000607626 Vibrio cholerae Species 0.000 description 1
- 241000607734 Yersinia <bacteria> Species 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000025845 adhesion to host Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 229940124350 antibacterial drug Drugs 0.000 description 1
- 244000000005 bacterial plant pathogen Species 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 238000003390 bioluminescence detection Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 230000021164 cell adhesion Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000006059 cover glass Substances 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- 208000006278 hypochromic anemia Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000009545 invasion Effects 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 244000000003 plant pathogen Species 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 238000007430 reference method Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- QJVXKWHHAMZTBY-GCPOEHJPSA-N syringin Chemical compound COC1=CC(\C=C\CO)=CC(OC)=C1O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 QJVXKWHHAMZTBY-GCPOEHJPSA-N 0.000 description 1
- QJVXKWHHAMZTBY-KSXIZUIISA-N syringin Natural products COc1cc(C=CCO)cc(OC)c1O[C@H]2O[C@@H](CO)[C@H](O)[C@@H](O)[C@@H]2O QJVXKWHHAMZTBY-KSXIZUIISA-N 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 229940118696 vibrio cholerae Drugs 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/142—Amino acids; Derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/20—Inorganic substances, e.g. oligoelements
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/015—Inorganic compounds
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/03—Organic compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
- A61K31/198—Alpha-amino acids, e.g. alanine or edetic acid [EDTA]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/025—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/21—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Pseudomonadaceae (F)
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
- G01N2333/24—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Enterobacteriaceae (F), e.g. Citrobacter, Serratia, Proteus, Providencia, Morganella, Yersinia
- G01N2333/245—Escherichia (G)
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Polymers & Plastics (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Inorganic Chemistry (AREA)
- Epidemiology (AREA)
- Environmental Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pest Control & Pesticides (AREA)
- Ecology (AREA)
- Nutrition Science (AREA)
- Biodiversity & Conservation Biology (AREA)
- Forests & Forestry (AREA)
- Dentistry (AREA)
- Plant Pathology (AREA)
- Animal Husbandry (AREA)
- Agronomy & Crop Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Botany (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
Abstract
氮源用于抑制细菌三型分泌系统的应用。本发明涉及生物医药领域,具体涉及氮源在制备用于预防和/或治疗病原菌侵染的药物中的应用。
Description
技术领域
本发明涉及生物医药领域,具体涉及氮源在制备用于预防和/或治疗病原菌侵染的药物中的应用。
背景技术
细菌感染一直是人类健康的重大威胁,虽然上世纪抗生素的发现为该问题提供了一个解决方案。但是由于耐药细菌等原因,我们需要寻找新的有效抗菌/抗感染方法。
抗生素能够杀死或抑制细菌的生长,近几十年来,被广泛应用于临床抗感染第一线。然而近年来,抗生素滥用、超级抗药细菌等原因,使得抗生素的杀菌/抑菌效率一降再降,因此,新型抗菌药物的研发是当前医学微生物研究领域的一个热点。微生物(特别是细菌)在长期进化中形成了不同于多细胞生物的独特繁殖优势,易对抗生素等生存压力产生耐药性(Drug resistance,在面临生存压力时仍能生长)或耐受性(Tolerance,在面临生存压力时不能生长但也不死亡),并使这种耐药或耐受能力在种群内(甚至不同物种间)进行传播。抗生素的不合理使用乃至滥用,加剧了致病菌耐药性的形成、并在世界范围内迅速蔓延,导致许多传染性疾病濒临无药可治的困境,给人类健康造成严重威胁。因此,如何抑制病原菌产生耐药,有效减少细菌感染,是生命医学领域面临的一个巨大挑战。抗生素在临床、农业及养殖业的大量使用乃至滥用,对药物敏感菌施加了生存选择压力,引起耐药菌群不断富集、倍增,最终导致耐药能力的产生和传播。
抗生素耐药的经典应对策略是不断研发新型抗生素或已有抗生素的新衍生物,以此治疗已对旧抗生素耐药的病原菌感染。该策略在过去半个世纪硕果累累,但目前已面临下述严峻挑战:(1)绝大多数抗生素靶点已经被研究和开发,发现有效的新型靶点已经非常困难,使得开拓全新抗生素举步维艰;(2)几乎所有已知抗生素母体化合物都已被变构优化,找到更高活性新衍生物的机会越来越小;(3)抗生素用药周期短,利润相对不高,而临床毒副作用审批要求严格,使得大多数国际知名制药厂商因商业利益解散了其抗生素研发团队,导致过去20多年罕有新抗生素投放市场;(4)过去十几年里,基因组、转录组、蛋白质组、代谢组等“OMICS”技术曾带来很大希望,认为对细菌“OMICS”的深入理解会提供无数的抗生素新靶点,从而开发出多种新抗生素;不幸的是,该平台技术耗费了巨大的人力物力,目前却连一个新抗生素都没能开发上市。
此外,临床上使用抑菌类抗生素时,只能抑制细菌繁殖,而无法有效清除细菌;使用杀菌类抗生素时,病人体内革兰氏阴性菌被杀死裂解后,往往会导致内毒素及其他致病物质在短时间内大量释放,从而可能加重患者病情。因此,抗生素耐药问题很难通过不断研发新抗生素的方法解决,急需科学家探索和开发不依赖抗生素的新策略。
三型分泌系统(Type 3 secretion system,T3SS)广泛存在于革兰氏阴性细菌,主要由包含5个操纵子(LEE1/2/3/4/5)的LEE毒力岛编码。其在细菌膜上形成的一种蛋白复合体跨膜结构,负责分泌蛋白因子,或将其直接注入高等宿主细胞,从而改变细胞的正常行为,导致宿主致病。T3SS与其他黏附因子类似,是细菌攻击宿主细胞的一种入侵武器,但其分子结构非常精密复杂,在细菌对宿主的附着和侵入过程中起着关键作用。T3SS在常见的革兰氏阴性致病菌中均有发现,如肠致病/出血性大肠杆菌(EnteropathogenicEscherichia coli—EPEC/Enterohemorrhagic Escherichia coli—EHEC)、鼠柠檬酸杆菌(Citrobacter rodentium—CR)、志贺氏菌、沙门氏菌、耶尔氏杆菌、假单胞菌、衣原体、霍乱弧菌、克雷伯氏菌和伯克氏菌等。已有研究表明,T3SS的功能缺失或其表达发生下降,会极大削弱病原菌对宿主的侵染致病能力。因此,探索病原微生物T3SS的作用机理,对于发现有效控制和治疗病原菌的新思路具有重要意义。
鉴于细菌T3SS在宿主侵染中起到的关键作用。近年来,世界各地的科学家尝试通过抑制T3SS表达,从而降低病原菌的侵染力。其先期结果显示了一定的应用前景。
发明内容
本发明第一方面提供一种抑制包含三型分泌系统的病原菌对宿主侵染的方法,所述方法包括:使所述病原菌接触有效量的氮源。
在本发明的一个实施方案中,所述方法通过抑制病原菌对宿主的定殖能力以达到抑制病原菌对宿主的侵染。
在本发明的一个实施方案中,所述方法通过抑制三型分泌系统相关蛋白的表达及其转运以达到抑制病原菌对宿主的定殖。
本发明第二方面提供氮源在制备用于预防和/或治疗包含三型分泌系统的病原菌侵染的药物中的应用。
本发明第三方面提供氮源作为病原菌三型分泌系统抑制剂的应用。
本发明第四方面在于提供一种病原菌三型分泌系统抑制剂。
本发明第五方面在于提供氮源在生鲜食品加工/清洗中的应用。
在本发明的一个实施方案中,所述病原菌是包含三型分泌系统的细菌,例如,所述病原菌可以是包含三型分泌系统的人畜共患病原菌、动物病原菌和植物病原菌。
在本发明的一个具体实施方案中,所述细菌属于选自下述的细菌属:肠致病/出血性大肠杆菌(Enteropathogenic Escherichia coli—EPEC/EnterohemorrhagicEscherichia coli—EHEC)、鼠柠檬酸杆菌(Citrobacter rodentium—CR)和丁香假单胞菌(Pseudomonas syringe)。
在本发明的一个实施方案中,所述宿主可以是人、动物和植物以及所有依赖于细菌三型分泌系统可以粘附定殖的表面。
在本发明的一个实施方案中,所述氮源选自无机氮源和有机氮源。
在本发明的一个实施方案中,所述氮源是氨基酸氮源。
在本发明的一个实施方案中,所述氮源选自谷氨酰胺、谷氨酸、天冬酰胺和天冬氨酸。
在本发明的一个实施方案中,所述氮源是谷氨酰胺。
在本发明的一个实施方案中,所述无机氮源选自铵盐和硝酸盐。
发明的有益效果
本发明通过添加氮源(特定氨基酸氮源/无机含氮化合物)作为环境信号,抑制病原菌中三型分泌系统相关蛋白的表达及其转运,减少其在宿主体内的定殖,促进病原菌排出,从而降低甚至消除病原菌感染风险。并且,促进病原菌排出过程中不伤害其本身,因此能够避免因生存压力而产生的耐药性,能够针对性的对抗目前因抗生素滥用而产生的大量耐药细菌,也对一般的细菌病害具有广谱作用。
本发明在细菌体外培养中实现了氮源(特定氨基酸氮源/无机含氮化合物)对于细菌毒力相关蛋白表达及其转运的抑制(约10-20倍),减少在细胞上黏附作用(约10倍),促进病原菌从小鼠体内的排出(100-1000倍),降低病原菌在小鼠肠道的定殖量。
附图说明
图1经过谷氨酰胺处理与未经谷氨酰胺处理的出血性大肠杆菌(EHEC)的生长情况。
图2谷氨酰胺对EHEC三型分泌系统蛋白分泌的抑制,其中图A为在培养基中添加水(对照)和谷氨酰胺后蛋白EspA/B/D的SDS-PAGE分析;图B为在培养基中添加水(对照)和谷氨酰胺后蛋白EspD/B的分泌量。
图3 EHEC中操纵子LEE4和LEE5的表达量。
图4 EHEC对宿主细胞的黏附,其中图A为荧光标记的EHEC和DAPI染色的细胞核的荧光显微镜下图;图B为EHEC对宿主细胞黏附情况的计数分析。
图5 CR分泌蛋白的SDS-PAGE分析。
图6在饮水中添加谷氨酰胺的实验组与未添加谷氨酰胺的对照组小鼠粪便中CR数量的统计。
图7在饮水中添加谷氨酰胺的实验组与未添加谷氨酰胺的对照组小鼠肠道中CR数量统计,其中图A为小鼠肠道中荧光素标记的CR检测分析;图B为小鼠肠道中荧光素标记的CR数量统计分析,P值<0.05。
图8在饮水中添加谷氨酰胺的实验组与未添加谷氨酰胺的对照组tlr4突变小鼠感染CR后存活率数量统计。
图9在饮水中添加谷氨酰胺的实验组与未添加谷氨酰胺的对照组小鼠粪便中EHEC数量的统计。
图10丁香假单胞菌(Pseudomonas syringae)毒力基因avrE,avrptoB和hrpL的mRNA表达量。
图11丁香假单胞菌(Pseudomonas syringae)感染拟南芥叶片致病实验结果。
图12经过不同氮源处理与未经处理的出血性大肠杆菌(EHEC)的生长情况(图B)及其对EHEC三型分泌系统蛋白分泌的抑制(图A)。
具体实施方式
下面将结合实施例对本发明的实施方案进行详细描述,但是本领域技术人员将会理解,下列实施例仅用于说明本发明,而不应视为限定本发明的范围。实施例中未注明具体条件者,按照常规条件或制造商建议的条件进行。所用试剂或仪器未注明生产厂商者,均为可以通过市购获得的常规产品。
MEM-Hepes培养基购自Sigma-Aldrich公司,货号M7278。所用的仪器IVIS luminaII购自Caliper Life Sciences公司。
实施例1谷氨酰胺抑制三型分泌系统转运蛋白分泌而不影响细菌生长
1.挑取出血性大肠杆菌(EHEC)单菌落,接种至LB肉汤培养基中,37℃,200rpm培养过夜;
2.将菌液通过4℃,4500xg离心30mins收集得到菌体和培养上清液;
3.将离心得到的菌体用MEM-Hepes培养基重悬后,按初始OD600=0.05重新接种到新鲜MEM-Hepes培养基中(分别添加终浓度为2mM的谷氨酰胺/谷氨酸/无机氮源,或相应体积无菌水),培养至OD600=0.6后,通过4℃,4500x g离心30mins收集得到菌体和培养上清液;
4.培养上清液通过0.45μm滤器过滤后加入TCA(终浓度10%),4℃过夜沉淀后,通过4℃,4500x g离心30mins后,弃去上清,用0.5M Tris-Cl(pH8.6)缓冲液重悬沉淀得到分泌蛋白样品;
5.将菌体重悬于PBS缓冲液中,超声破碎菌体后,通过4℃,4500x g离心30mins收集上清得到菌体蛋白裂解样品;
我们对于EHEC的分泌蛋白进行SDS-PAGE分析,实验结果表明应用氮源(谷氨酰胺/谷氨酸/无机氮源)处理后不抑制EHEC生长(图1,图12B),但其三型分泌系统转运蛋白输出减少了约20倍(图2,图12A)。
报告基因实验结果(图3)表明应用氮源(谷氨酰胺)处理后EHEC的LEE4和LEE5操纵子的基因表达量也下降了10-20倍。
细菌的生长曲线测定:
将细菌按以上培养方法(步骤1、2、3)分别在添加不同氮源的培养基中培养,在设定时间点分别取出1ml菌液加入比色皿,通过分光光度计测量OD600值。
LEE4/5表达水平的测定:(参考文献:Analysis of the expression,regulationand export of NleA-E in Escherichia coli O157:H7.Microbiology.2007)
参考文献方法,将LEE4-gfp/LEE5-gfp质粒分别通过电穿孔方法转入待测菌中,在相应抗性平板上挑取转化菌落,转化菌落按以上培养方法在添加不同氮源及抗生素的培养基中培养,当菌液OD600值到达0.8时,取150μl菌液在多功能酶标仪中读取绿色荧光值,即为LEE4/5的相对表达值。
实施例2细胞黏附实验
1.将Hela细胞接种到腔室培养玻片中,37℃,5%CO2培养至细胞密度>80%;
2.挑取EHEC单菌落,接种至LB肉汤培养基中,37℃,200rpm培养过夜;
3.将过夜培养的菌液1:100稀释到新鲜MEM-Hepes培养基中(添加终浓度为2mM的谷氨酰胺或相应体积无菌水),37℃,200rpm培养至OD600=0.6;
4.按M.O.I.1:100将EHEC加入Hela细胞培养基(添加终浓度为2mM的谷氨酰胺或相应体积无菌水)中,37℃,5%CO2孵育1hr,弃去培养基,加入4%PFA室温固定30mins;
5.弃去4%PFA,加入6%BSA孵育1小时后弃去,加入FITC标记的O157抗体孵育1小时用于标记EHEC,弃去O157抗体后加入PBS清洗3遍,弃去PBS;
6.去除玻片上的腔室,加入适量含有DAPI染液的封片液,盖上盖玻片,置于暗盒内,4℃,24hrs固化;
7.荧光显微镜下对荧光标记的EHEC及DAPI染色的细胞核进行观察计数。
从图4中可以看出,在不添加谷氨酰胺的对照组中,EHEC能在Hela细胞上形成有效黏附,从而为进一步感染细胞奠定基础。而在添加谷氨酰胺的实验组中,EHEC在Hela细胞上形成有效黏附的能力大大下降,很难造成有效侵染。通过计数分析证实,添加谷氨酰胺大大降低了EHEC对于Hela细胞的黏附能力,黏附数量仅为不添加谷氨酰胺组的10%左右甚至更低。
实施例3动物感染实验
1.选取20只6周龄BALB/C小鼠(CR感染致死实验使用B10ScNJ小鼠),分为对照组(n=10)和实验组(n=10),对照组小鼠的饮食中不含谷氨酰胺/谷氨酸,实验组小鼠额外在饮水中添加谷氨酰胺;
2.挑取转入荧光质粒(pGEN-luxCDABE)的病原菌单菌落,接种至LB肉汤培养基中,37℃,200rpm培养过夜;
3.离心获得过夜菌,加入PBS清洗3遍后以1x109菌量对小鼠进行灌胃(灌胃前对小鼠进行24小时断水断食);
4.收集小鼠粪便,加入生理盐水匀浆后,进行10倍梯度稀释点板计数分析;
5.第9天和第15天,每组分别随机处死5只小鼠,进行解剖,取出肠道,使用IVISLumina II进行生物发光检测计数分析。
CR动物感染实验:从图中可以看出,添加谷氨酰胺会使得体外培养CR上清中的分泌蛋白减少(图5),且在CR感染小鼠饮食中添加谷氨酰胺后,会导致该病原菌的大量排出(图6、9),从而导致在小鼠肠道中的CR大大减少(图7)。在tlr4突变的B10ScNJ小鼠(CR小鼠感染致死模型)中使用CR进行感染时,CR感染对照组小鼠在感染23天后50%(5只)死亡,而饮食添加谷氨酰胺的小鼠在感染23天后仅有10%(1只)死亡(图8)。饮食添加谷氨酰胺对于CR感染致死小鼠表现出良好的保护效果。
EHEC动物感染实验:在EHEC感染小鼠模型中,也可观察到类似于CR感染小鼠模型中的结果,在感染小鼠饮食中添加谷氨酰胺后,会导致EHEC的大量排出(图9),由于EHEC在小鼠体内感染定殖量不如CR高,无法用生物发光实时检测EHEC在肠道内定殖情况。
实施例4植物病原菌毒力基因的mRNA表达水平检测
1.从平板上挑取丁香假单胞菌番茄致病株(DC3000)单菌落,将细菌接种于液体KB培养基,置24℃250rpm摇床培养至对数中期,再转移至MM培养基,其中实验组按照1:100的比例添加200mM L-谷氨酰胺溶液(终浓度2mM L-谷氨酰胺),对照组添加相同体积的无菌水,于250rpm,24℃条件下培养12h后通过4℃,4600xg离心得到菌体。
2.细菌RNA提取:参照Roche Tripure试剂盒说明书提供的方法对步骤1离心得到的菌体进行操作。
3.反转录反应:参照abcam的5×All In One RT MasterMix试剂盒说明书提供的方法对步骤2得到的细菌RNA进行反转录操作。
4.RT-PCR(或qPCR)反应体系和反应程序参照Takara的Premix Ex TaqII(Tli RNaseH Plus)试剂说明书,引物见下表。反应结束后,先观察Real Time PCR的扩增曲线和融解曲线是否正确,并根据得到的CT值进行分析作图。
RT-PCR引物信息
引物名称 | 序列 | SEQ ID |
avrPtoB-FP | TGCACACGCCAACAGTATCG | SEQ ID NO:1 |
avrPtoB-RP | CGCAGGTGCTCTAAATTGAC | SEQ ID NO:2 |
avrE-FP | ACTTGTCGAGCCCGTTCATG | SEQ ID NO:3 |
avrE-RP | GTCAGGCCGCGATTGTTGTTC | SEQ ID NO:4 |
hrpL-FP | CATCTTCGTCCGCCGGTATTC | SEQ ID NO:5 |
hrpL-RP | GTCCATCTCCAGCGACACTTC | SEQ ID NO:6 |
psy-16s-FP | CGGTAATACAGAGGGTGCAAGC | SEQ ID NO:7 |
psy-16s-RP | ATTCCACCACCCTCTACCATAC | SEQ ID NO:8 |
从实验结果来看,丁香假单胞菌在添加谷氨酰胺后,与对照组相比,其三型分泌系统相关的毒力基因的mRNA表达水平明显减少(降低量>60%)(图10),该病原菌的毒力水平明显下调。
实施例5植物病原菌致病实验
用108CFU/mL(OD600≈0.2)的丁香假单胞菌番茄致病株(Pseudomonas syringae)DC3000细菌悬浮液通过注射的方式感染野生型拟南芥,会引起植物宿主的缓慢的萎黄症状。因此,在该实验中我们选择这一剂量对植株进行感染。
1.从平板上挑取丁香假单胞菌番茄致病株(DC3000)单菌落,将细菌接种于液体KB培养基,置24℃250rpm摇床培养至对数中期,再转移至MM培养基,其中实验组按照1:100的比例添加200mM L-谷氨酰胺溶液(终浓度2mM L-谷氨酰胺),对照组添加相同体积的无菌水,于250rpm,24℃条件下培养12h后通过4℃,4600xg离心得到菌体。
2.离心后菌体最终用10mM氯化镁溶液重悬,使菌液浓度为OD600=0.2,同时加入Silwet L-77(终体积分数为0.02%)。选取4周龄的野生型拟南芥(Col-0),将菌悬液通过注射方式感染叶片上。感染后观察叶片颜色、形态的变化。
从图11中可以观察到,感染后第4天,对照组(无菌水)的叶片出现了明显的黄色斑点,且在第7天,对照组(无菌水)叶片的黄色区域已明显扩散开来,并出现枯萎现象,而实验组(2mM L-谷氨酰胺)的叶片在感染后第4天或第7天所出现的黄色斑点较对照组明显更少,外在表征更为健康。
以上对本发明做了详尽的描述,其目的在于让本领域技术人员能够了解本发明的内容并加以实施,并不能以此限制本发明的保护范围,凡根据本发明的精神实质所做的等效变化或修饰都应涵盖在本发明的范围内。
序列表
<110> 厦门大学
<120> 氮源用于抑制细菌三型分泌系统的应用
<130> IDC190218
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 20
<212> DNA
<213> 人工序列
<400> 1
tgcacacgcc aacagtatcg 20
<210> 2
<211> 20
<212> DNA
<213> 人工序列
<400> 2
cgcaggtgct ctaaattgac 20
<210> 3
<211> 20
<212> DNA
<213> 人工序列
<400> 3
acttgtcgag cccgttcatg 20
<210> 4
<211> 21
<212> DNA
<213> 人工序列
<400> 4
gtcaggccgc gattgttgtt c 21
<210> 5
<211> 21
<212> DNA
<213> 人工序列
<400> 5
catcttcgtc cgccggtatt c 21
<210> 6
<211> 21
<212> DNA
<213> 人工序列
<400> 6
gtccatctcc agcgacactt c 21
<210> 7
<211> 22
<212> DNA
<213> 人工序列
<400> 7
cggtaataca gagggtgcaa gc 22
<210> 8
<211> 22
<212> DNA
<213> 人工序列
<400> 8
attccaccac cctctaccat ac 22
Claims (10)
1.一种抑制包含三型分泌系统的病原菌对宿主侵染的方法,所述方法包括:使所述病原菌接触有效量的氮源。
2.根据权利要求1所述的方法,其中所述氮源通过抑制病原菌对宿主的定殖能力以减少病原菌对宿主的侵染和促进病原菌从宿主体内的排出。
3.根据权利要求1或2所述的方法,其中所述氮源抑制三型分泌系统毒力相关蛋白的表达及其转运。
4.氮源在制备用于预防和/或治疗包含三型分泌系统的病原菌侵染的药物中的用途。
5.氮源作为病原菌三型分泌系统抑制剂的用途。
6.根据权利要求1-3任一项所述的方法或权利要求4-5任一项所述的用途,其中所述病原菌是革兰氏阴性细菌,优选地,所述病原菌选自:肠致病/出血性大肠杆菌(Enteropathogenic Escherichia coli—EPEC/Enterohemorrhagic Escherichia coli—EHEC)、鼠柠檬酸杆菌(Citrobacter rodentium—CR)和丁香假单胞菌(Pseudomonassyringae)。
7.根据权利要求1-3任一项所述的方法或权利要求4-5任一项所述的用途,其中所述宿主是人、动物和植物。
8.根据权利要求1-3任一项所述的方法或权利要求4-5任一项所述的用途,其中所述氮源选自无机氮源和有机氮源,优选地所述有机氮源选自谷氨酰胺、谷氨酸、天冬酰胺和天冬氨酸;所述无机氮源选自铵盐和硝酸盐。
9.一种治疗革兰氏阴性致病菌感染的药物,其特征在于:所述药物含有权利要求8所定义的氮源。
10.一种治疗革兰氏阴性致病菌感染的食品或饲料添加剂或清洗剂,其特征在于:所述添加剂或清洗剂含有权利要求8所定义的氮源。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010631326.XA CN113876952B (zh) | 2020-07-03 | 2020-07-03 | 氮源用于抑制细菌三型分泌系统的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202010631326.XA CN113876952B (zh) | 2020-07-03 | 2020-07-03 | 氮源用于抑制细菌三型分泌系统的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113876952A true CN113876952A (zh) | 2022-01-04 |
CN113876952B CN113876952B (zh) | 2023-04-14 |
Family
ID=79013195
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202010631326.XA Active CN113876952B (zh) | 2020-07-03 | 2020-07-03 | 氮源用于抑制细菌三型分泌系统的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113876952B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1878788A (zh) * | 2004-02-09 | 2006-12-13 | 社团法人北里研究所 | K01-0509物质和其生产方法 |
CN102524573A (zh) * | 2012-01-06 | 2012-07-04 | 四川农业大学 | 一种建鲤饲料添加剂 |
CN102871996A (zh) * | 2012-09-10 | 2013-01-16 | 中国医学科学院医药生物技术研究所 | 一种抗菌药物组合物及其应用 |
US20180187238A1 (en) * | 2015-06-26 | 2018-07-05 | Saber Biotics, Llc | Selective media and uses thereof |
-
2020
- 2020-07-03 CN CN202010631326.XA patent/CN113876952B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1878788A (zh) * | 2004-02-09 | 2006-12-13 | 社团法人北里研究所 | K01-0509物质和其生产方法 |
CN102524573A (zh) * | 2012-01-06 | 2012-07-04 | 四川农业大学 | 一种建鲤饲料添加剂 |
CN102871996A (zh) * | 2012-09-10 | 2013-01-16 | 中国医学科学院医药生物技术研究所 | 一种抗菌药物组合物及其应用 |
US20180187238A1 (en) * | 2015-06-26 | 2018-07-05 | Saber Biotics, Llc | Selective media and uses thereof |
Non-Patent Citations (1)
Title |
---|
许志刚: "《普通植物病理学》", 31 July 2009 * |
Also Published As
Publication number | Publication date |
---|---|
CN113876952B (zh) | 2023-04-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Chen et al. | The quorum quenching bacterium Bacillus licheniformis T-1 protects zebrafish against Aeromonas hydrophila infection | |
JP2022106717A (ja) | 微生物集団の改変および微生物相の修飾 | |
US20230193241A1 (en) | Altering microbial populations & modifying microbiota | |
CN109735471B (zh) | 一株微小杆菌及其作为益生菌在水产上的应用 | |
US8821855B2 (en) | Methods for isolating phage and for controlling microorganism populations with the phage | |
CN113040390B (zh) | 一株益生、耐盐约氏乳杆菌及其在畜禽水产养殖中防治病原菌的应用 | |
EP2356991B1 (en) | Use of bacteria of the genus tenacibaculum for quorum quenching | |
Akbari Kiarood et al. | Quorum‐quenching endophytic bacteria inhibit disease caused by Pseudomonas syringae pv. syringae in Citrus cultivars | |
KR20220019697A (ko) | 항박테리아제 및 방법 | |
Leister et al. | Genome analysis of Enterobacter asburiae and Lelliottia spp. proliferating in oligotrophic drinking water reservoirs and lakes | |
CN113876952B (zh) | 氮源用于抑制细菌三型分泌系统的应用 | |
CN102027107A (zh) | 减少细菌中接合型质粒 | |
Cheng et al. | Dual RNA sequencing analysis of Bacillus amyloliquefaciens and Sclerotinia sclerotiorum during infection of soybean seedlings by S. sclerotiorum unveils antagonistic interactions | |
Gao et al. | Transcriptomic and phenotype analysis revealed the role of rpoS in stress resistance and virulence of pathogenic Enterobacter cloacae from Macrobrachium rosenbergii | |
Hayashi et al. | Cas9-assisted biological containment of a genetically engineered human commensal bacterium and genetic elements | |
CN107904187A (zh) | 对迟缓爱德华氏菌具有高拮抗作用的融合魏斯氏菌3dw | |
Suresh et al. | Predatory efficacy of Bdellovibrio stolpii isolated from the wastewater sources against the multidrug-resistant clinical isolates | |
CN114736834B (zh) | 一种短小芽孢杆菌ts1及其应用 | |
Ozdemir | Curing the drug resistance plasmid in E. coli O157: H7. | |
Gavu | Taxonomy, spoilage, and virulence characteristics of Kaistella species isolated from fish | |
Zaima | A study on relationship of Plasmids with Azithromycin and Ciprofloxacin Resistance in isolates causing Urinary Tract Infection | |
Walker-Sünderhauf | Removal of antimicrobial resistance genes from bacterial strains and communities using CRISPR-Cas9 | |
CN116716261A (zh) | 一株不易产生抗性的蜡样芽孢杆菌噬菌体dz1及其应用 | |
Hossain et al. | Bacteriophage and non-pathogenic vibrio to control diseases in black tiger shrimp (Penaeus monodon) aquaculture | |
KR20230090858A (ko) | 붉바리 치어 양식용 프로바이오틱스 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |