CN107012109A - Applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed - Google Patents
Applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed Download PDFInfo
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- CN107012109A CN107012109A CN201710211207.7A CN201710211207A CN107012109A CN 107012109 A CN107012109 A CN 107012109A CN 201710211207 A CN201710211207 A CN 201710211207A CN 107012109 A CN107012109 A CN 107012109A
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- escherichia coli
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- phage infection
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- 230000011664 signaling Effects 0.000 title claims abstract description 26
- 241000588724 Escherichia coli Species 0.000 title claims abstract description 21
- 208000015181 infectious disease Diseases 0.000 title claims abstract description 16
- 238000011534 incubation Methods 0.000 claims abstract description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 7
- 210000002429 large intestine Anatomy 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 6
- 241001013691 Escherichia coli BW25113 Species 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 description 2
- YRYOXRMDHALAFL-UHFFFAOYSA-N N-(3-oxohexanoyl)homoserine lactone Chemical compound CCCC(=O)CC(=O)NC1CCOC1=O YRYOXRMDHALAFL-UHFFFAOYSA-N 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 229940041514 candida albicans extract Drugs 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 241001515965 unidentified phage Species 0.000 description 2
- 239000012138 yeast extract Substances 0.000 description 2
- UKAUYVFTDYCKQA-UHFFFAOYSA-N -2-Amino-4-hydroxybutanoic acid Natural products OC(=O)C(N)CCO UKAUYVFTDYCKQA-UHFFFAOYSA-N 0.000 description 1
- -1 Acyl homoserine lactones Chemical class 0.000 description 1
- NLXLAEXVIDQMFP-UHFFFAOYSA-N Ammonia chloride Chemical compound [NH4+].[Cl-] NLXLAEXVIDQMFP-UHFFFAOYSA-N 0.000 description 1
- 108091033409 CRISPR Proteins 0.000 description 1
- 238000010354 CRISPR gene editing Methods 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 102000006465 DNA Restriction-Modification Enzymes Human genes 0.000 description 1
- 108010044289 DNA Restriction-Modification Enzymes Proteins 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000032770 biofilm formation Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
- 229910000397 disodium phosphate Inorganic materials 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 125000003104 hexanoyl group Chemical group O=C([*])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- VLSMHEGGTFMBBZ-OOZYFLPDSA-N kainic acid Chemical compound CC(=C)[C@H]1CN[C@H](C(O)=O)[C@H]1CC(O)=O VLSMHEGGTFMBBZ-OOZYFLPDSA-N 0.000 description 1
- 229910052943 magnesium sulfate Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000018612 quorum sensing Effects 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000003014 reinforcing effect Effects 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- 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
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Tropical Medicine & Parasitology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed.By taking ETEC BW25113, DH5 α and BL21 as an example, by adding signaling molecule C6 AHL co-incubations, greatly reduce plaque number, it was demonstrated that signaling molecule C6 AHL have the effect suppressed in Phage Infection Escherichia coli.
Description
Technical field
The present invention relates to applications of the signaling molecule C6-AHL in Phage Infection Escherichia coli are suppressed, belong to microorganism neck
Domain.
Background technology
Escherichia coli are prokaryotes, construct relatively easy, and genetic background is clear, and culture operation is easy, thus usually by
It is used as the object of genetic engineering, or cytologic experiment is participated in as model organism.But in application process, large intestine bar
Bacterium usually can be by Phage Infection.At present frequently with absorption suppress, injection retardance, abortive infection, restriction modification system,
The methods such as CRISPR systems prevent Phage Infection Escherichia coli, but the above method all relies on bacterium its own system and play work
With the growth metabolism situation of its effect and bacterium is closely related, and influences the factor of bacterial growth a lot, to final effect band
Carry out many uncertain obstructions.
Acyl homoserine lactones（AHL）It is the signaling molecule that a class mediates gram negative bacteria quorum sensing, can passes through
Free diffusing is entered in microbial cell, and acceptor corresponding with intracellular is combined, so as to activate or suppress the expression of target gene, often
Studied for biofilm formation, cell mobility etc., for example：It is added in bio-denitrifying sewage system, reinforcing nitrifier is biological
The formation of film, so as to help to be enriched with nitrifier, improves nitrification efficiency.AHL signaling molecules are suppressing Phage Infection large intestine bar
Application in bacterium does not have been reported that also.
The content of the invention
It is an object of the invention to provide signaling molecule C6-AHL suppress Phage Infection Escherichia coli in application, specifically
Step is：Signaling molecule C6-AHL is dissolved in DMSO solution is made, added in M9 minimal mediums, it is 5 μ to make C6-AHL concentration
M, in 33 DEG C, Escherichia coli OD600With Escherichia coli co-incubation under conditions of=1.2.
Signaling molecule C6-AHL, chemical name is N- (β -one hexanoyl)-DL- homoserine lactones（N-(β-
Ketocaproyl)-DL- homoserine lactone）, No. CAS is 76924-95-3, and molecular formula is C10H15NO4, structural formula
For：
Beneficial effects of the present invention：
（1）The present invention can effectively suppress Phage Infection Escherichia coli（T4 Phage Infection Escherichia coli BW25113, DH5 α,
Downward trend is presented in BL21 ability）；
（2）The effect that the present invention suppresses Phage Infection Escherichia coli is not influenceed by the upgrowth situation of Escherichia coli, effect stability
It is controllable.Escherichia coli do not produce C6-AHL signaling molecules in itself, but with the ability for perceiving and combining C6-AHL signaling molecules
（Escherichia coli pass through a receptor protein in cytoplasm（SdiA）Carry out perceptual signal molecule C6-AHL and make it into signal
In path, so as to play the effect of correlation, the expression quantity of the albumen is relatively stable within whole cell life cycle and deposits always
）, C6-AHL signaling molecules are added by external source, the amount of signaling molecule is easily controllable, signaling molecule quantifies, not by Escherichia coli
Upgrowth situation influence；
（3）Pollution of the present invention to preventing and treating biological products fermentation process pnagus medius to Escherichia coli has great importance.
Brief description of the drawings
Fig. 1 is ETEC BW25113 plaques in the case of without signaling molecule C6-AHL in embodiment 1
Growing state；
Fig. 2 is the growth of ETEC BW25113 plaques in the case where adding signaling molecule C6-AHL in embodiment 1
Situation；
Fig. 3 is that embodiment 1 to 3 Escherichia coli BW25113, DH5 α, BL21 infects without with addition C6-AHL signaling molecules T4
Rate（That is plaque growth number percentage）.
Embodiment
The essentiality content of the present invention is further illustrated with reference to the accompanying drawings and examples, but this hair is not limited with this
It is bright.The reagent and instrument used in embodiment, is conventional reagent and conventional method unless otherwise specified.
Signaling molecule C6-AHL：Purchased from Sigma companies, production code member is K3255.
M9 minimal mediums are formulated：200mL 5 × M9salts, 1mL 1M MgSO4, the glucose of 10mL 20%, 0.1mL
1M CaCl2, aqua sterilisa to 1L.5 × M9salts wherein relative to every liter, it contains：64g Na2HPO4×7H2O, 15g
KH2PO4, 2.5g NaCl, 5g NH4Cl。
LB solid culture based formulas：Tryptone（Tryptone）10g, Yeast Extract（Yeast extract）5g,
NaCl 10g, Agar（Agar）15g, adds distilled water to 1L.121 DEG C of sterilizing 20min.
Embodiment 1
Meet ETEC BW25113（CICC 23872）In 5mL M9 culture mediums, 33 DEG C, 150rpm incubated overnights, the
It is connected to 2% inoculum concentration in 30mL M9 culture mediums within two days, control group adds 17.5 μ L DMSO, experimental group adds 17.5 μ L
0.2g/100mL signaling molecule C6-AHL DMSO solution（It is now with the current）.OD is determined with ultraviolet specrophotometer600For 1.2.
Gradient dilution T4 bacteriophages, are diluted to 10-7, take 10-6、10-7T4 dilutions.300µL OD600=1.2 Escherichia coli BW25113
Bacterium solution and 100 μ L 10-6、10-7T4 dilutions are mixed, in 33 DEG C of incubator absorption 6min.Then detected with double-layer agar technique, often
Individual sample measure 3 is parallel.Good flat board solidifies in superclean bench, is then inverted to be placed in 33 DEG C of incubators and cultivates
3h, carries out plaque count.As shown in Figure 1, Figure 2 and Figure 3, control group plaque number is 289 to experimental result, and experimental group is bitten
Bacterial plaque number is 253, have dropped 12.5%, shows that external source addition signaling molecule C6-AHL can effectively suppress T4 bacteriophages and invade
Contaminate Escherichia coli BW25113.
Embodiment 2
Using ETEC DH5 α（CICC 10339）Tested, experimental method be the same as Example 1.Experimental result such as Fig. 3
Shown, control group plaque number is 109, and experimental group plaque number is 91, have dropped 16.5%, shows that external source is added
Signaling molecule C6-AHL can effectively suppress T4 Phage Infection bacillus coli DH 5 alphas.
Embodiment 3
Using ETEC BL21（CICC 23796）Tested, experimental method be the same as Example 1.Experimental result such as Fig. 3
Shown, control group plaque number is 305, and experimental group plaque number is 277, have dropped 9.2%, shows that external source is added
Signaling molecule C6-AHL can effectively suppress T4 Phage Infection e. coli bl21s.
Claims (2)
1. applications of the signaling molecule C6-AHL in Phage Infection Escherichia coli are suppressed.
2. application according to claim 1, is concretely comprised the following steps：Signaling molecule C6-AHL is dissolved in DMSO solution is made, added
Add in M9 minimal mediums, it is 5 μM to make C6-AHL concentration, in 33 DEG C, Escherichia coli OD600With large intestine bar under conditions of=1.2
Bacterium co-incubation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211207.7A CN107012109A (en) | 2017-04-01 | 2017-04-01 | Applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710211207.7A CN107012109A (en) | 2017-04-01 | 2017-04-01 | Applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed |
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| Publication Number | Publication Date |
|---|---|
| CN107012109A true CN107012109A (en) | 2017-08-04 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710211207.7A Pending CN107012109A (en) | 2017-04-01 | 2017-04-01 | Applications of the signaling molecule C6 AHL in Phage Infection Escherichia coli are suppressed |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108103031A (en) * | 2018-01-10 | 2018-06-01 | 浙江省淡水水产研究所 | A kind of wide range phage preparation used for aquiculture and preparation method thereof |
| CN109402072A (en) * | 2018-09-11 | 2019-03-01 | 昆明理工大学 | The purposes of signaling molecule C4-AHL |
| CN110129279A (en) * | 2019-04-24 | 2019-08-16 | 昆明理工大学 | A kind of enterococcus faecalis bacteriophage and its separation, purifying, enrichment and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105176932A (en) * | 2014-05-07 | 2015-12-23 | 因特伦生物技术株式会社 | Method for prevention and treatment of escherichia coli infections using a bacteriophage with broad antibacterial spectrum against escherichia coli |
| CN105504000A (en) * | 2016-01-21 | 2016-04-20 | 福建农林大学 | Method for screening N-acylhomoserine lactone simulant |
| WO2016164636A1 (en) * | 2015-04-09 | 2016-10-13 | The Regents Of The University Of California | Engineered bacteria for production and release of therapeutics |
-
2017
- 2017-04-01 CN CN201710211207.7A patent/CN107012109A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105176932A (en) * | 2014-05-07 | 2015-12-23 | 因特伦生物技术株式会社 | Method for prevention and treatment of escherichia coli infections using a bacteriophage with broad antibacterial spectrum against escherichia coli |
| WO2016164636A1 (en) * | 2015-04-09 | 2016-10-13 | The Regents Of The University Of California | Engineered bacteria for production and release of therapeutics |
| CN105504000A (en) * | 2016-01-21 | 2016-04-20 | 福建农林大学 | Method for screening N-acylhomoserine lactone simulant |
Non-Patent Citations (2)
| Title |
|---|
| HØYLAND-KROGHSBO ET AL.,: ""A Quorum-Sensing-Induced Bacteriophage Defense Mechanism"", 《MBIO》 * |
| 王楠等: ""群体感应及其介导的抗噬菌体研究进展"", 《中国微生态学杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108103031A (en) * | 2018-01-10 | 2018-06-01 | 浙江省淡水水产研究所 | A kind of wide range phage preparation used for aquiculture and preparation method thereof |
| CN108103031B (en) * | 2018-01-10 | 2021-09-07 | 浙江省淡水水产研究所 | Wide-spectrum phage preparation for aquaculture and preparation method thereof |
| CN109402072A (en) * | 2018-09-11 | 2019-03-01 | 昆明理工大学 | The purposes of signaling molecule C4-AHL |
| CN110129279A (en) * | 2019-04-24 | 2019-08-16 | 昆明理工大学 | A kind of enterococcus faecalis bacteriophage and its separation, purifying, enrichment and application |
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