Difference between revisions of "Part:BBa K3782008"
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This gene cluster is found naturally in <i>Pseudomonas syringae pv. aptata</i> DSM50252. It contains both structural genes, necessary to building the tailocin, and lysis genes, which lyse the producer cell in order to release the tailocins. The genes were identified using PHASTER, the detailed annotation results are available here<ref>PHASTER. https://phaster.ca/submissions/AEAN00000000.1</ref>. | This gene cluster is found naturally in <i>Pseudomonas syringae pv. aptata</i> DSM50252. It contains both structural genes, necessary to building the tailocin, and lysis genes, which lyse the producer cell in order to release the tailocins. The genes were identified using PHASTER, the detailed annotation results are available here<ref>PHASTER. https://phaster.ca/submissions/AEAN00000000.1</ref>. | ||
| − | [[File:T--UNILausanne--tailocin.png| | + | [[File:T--UNILausanne--tailocin.png|500px|thumb|center|'''Figure 1:''' Tailocin gene cluster of <i>Pseudomonas syringae</i> DSM50252. Gene annotation as identified by PHASTER<ref>PHASTER. https://phaster.ca/submissions/AEAN00000000.1</ref>.]]<br> |
This tailocin targets several strains of <i>Pseudomonas syringae</i>, a detailed list of which can be found in Baltrus et al. 2019<ref>Baltrus, D. A., Clark, M., Smith, C. & Hockett, K. L. Localized recombination drives diversification of killing spectra for phage-derived syringacins. ISME Journal 13, 237–249 (2019).</ref>. | This tailocin targets several strains of <i>Pseudomonas syringae</i>, a detailed list of which can be found in Baltrus et al. 2019<ref>Baltrus, D. A., Clark, M., Smith, C. & Hockett, K. L. Localized recombination drives diversification of killing spectra for phage-derived syringacins. ISME Journal 13, 237–249 (2019).</ref>. | ||
<br>In our project, we aimed at using this protein complex to kill the plant pathogen <i>Pseudomonas syringae pv. syringae</i> B301D. | <br>In our project, we aimed at using this protein complex to kill the plant pathogen <i>Pseudomonas syringae pv. syringae</i> B301D. | ||
| − | <br>We aimed at cloning it in <i>E. coli</i> BL21 pLys, in a vector carrying a lac inducible T7 promoter but were not able to achieve this due to lack of time. | + | <br><br>We aimed at cloning it in <i>E. coli</i> BL21 pLys, in a vector carrying a lac inducible T7 promoter but were not able to achieve this due to lack of time. |
| + | =Sequence and Features= | ||
| + | <partinfo>BBa_K3782008 SequenceAndFeatures</partinfo> | ||
| + | =References= | ||
| + | {{reflist}} | ||
Revision as of 15:50, 17 October 2021
Pseudomonas syringae DSM50252 derived tailocin gene cluster
Tailocin Gene Cluster
This part is the tailocin gene cluster of Pseudomonas syringae pv. aptata DSM50252. Tailocins are phage-derived bacteriocins produced by several species of bacteria in order to compete against closely related bacterial strains (often even of the same species) without harming cells from their producer’s strain[1]. They structurally look like phage tails and are encoded by phage genes repurposed by bacteria[2]. When tailocins encounter their target bacteria, they bind to their cell wall and contract, thus perforating the wall. This creates a pore, which dissipates the proton gradient and kill the target cell. This mechanism allows a potent killing efficiency, as only one or few particles are sufficient to kill the cell[3].
Profile
| Name | Tailocin Gene Cluster |
| Base pairs | |
| Number of amino acids | |
| Molecular weight | 25.35 kDa |
| Origin | Pseudomonas syringae pv. aptata DSM50252, extracted |
Usage and Biology
This gene cluster is found naturally in Pseudomonas syringae pv. aptata DSM50252. It contains both structural genes, necessary to building the tailocin, and lysis genes, which lyse the producer cell in order to release the tailocins. The genes were identified using PHASTER, the detailed annotation results are available here[4].
Figure 1: Tailocin gene cluster of Pseudomonas syringae DSM50252. Gene annotation as identified by PHASTER[5].
This tailocin targets several strains of Pseudomonas syringae, a detailed list of which can be found in Baltrus et al. 2019[6].
In our project, we aimed at using this protein complex to kill the plant pathogen Pseudomonas syringae pv. syringae B301D.
We aimed at cloning it in E. coli BL21 pLys, in a vector carrying a lac inducible T7 promoter but were not able to achieve this due to lack of time.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2030
Illegal EcoRI site found at 5264
Illegal EcoRI site found at 11011
Illegal PstI site found at 182
Illegal PstI site found at 606
Illegal PstI site found at 2085
Illegal PstI site found at 2348
Illegal PstI site found at 2387
Illegal PstI site found at 3804
Illegal PstI site found at 5597 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2030
Illegal EcoRI site found at 5264
Illegal EcoRI site found at 11011
Illegal PstI site found at 182
Illegal PstI site found at 606
Illegal PstI site found at 2085
Illegal PstI site found at 2348
Illegal PstI site found at 2387
Illegal PstI site found at 3804
Illegal PstI site found at 5597 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2030
Illegal EcoRI site found at 5264
Illegal EcoRI site found at 11011
Illegal BglII site found at 11166
Illegal BamHI site found at 1330
Illegal BamHI site found at 10503
Illegal XhoI site found at 9613 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2030
Illegal EcoRI site found at 5264
Illegal EcoRI site found at 11011
Illegal PstI site found at 182
Illegal PstI site found at 606
Illegal PstI site found at 2085
Illegal PstI site found at 2348
Illegal PstI site found at 2387
Illegal PstI site found at 3804
Illegal PstI site found at 5597 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2030
Illegal EcoRI site found at 5264
Illegal EcoRI site found at 11011
Illegal PstI site found at 182
Illegal PstI site found at 606
Illegal PstI site found at 2085
Illegal PstI site found at 2348
Illegal PstI site found at 2387
Illegal PstI site found at 3804
Illegal PstI site found at 5597
Illegal NgoMIV site found at 1365
Illegal NgoMIV site found at 1790
Illegal NgoMIV site found at 2072
Illegal NgoMIV site found at 2485
Illegal NgoMIV site found at 2747
Illegal NgoMIV site found at 3574
Illegal NgoMIV site found at 4028
Illegal AgeI site found at 1032
Illegal AgeI site found at 2377
Illegal AgeI site found at 3245
Illegal AgeI site found at 5176
Illegal AgeI site found at 6840
Illegal AgeI site found at 8051
Illegal AgeI site found at 9867 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 7258
Illegal BsaI site found at 8089
Illegal BsaI site found at 11930
Illegal BsaI site found at 12494
Illegal BsaI.rc site found at 1134
Illegal BsaI.rc site found at 3436
Illegal BsaI.rc site found at 3905
Illegal BsaI.rc site found at 4016
Illegal BsaI.rc site found at 7805
Illegal SapI.rc site found at 6651
References
Template:Reflist- ↑ Patz, S. et al. Phage tail-like particles are versatile bacterial nanomachines – A mini-review. Journal of Advanced Research vol. 19 75–84 (2019).
- ↑ Ghequire, M. G. K. & Mot, R. de. The Tailocin Tale: Peeling off Phage Tails. Trends in Microbiology 23, 587–590 (2015).
- ↑ Carim, S. et al. Systematic discovery of pseudomonad genetic factors involved in sensitivity to tailocins. ISME Journal (2021) doi:10.1038/s41396-021-00921-1.
- ↑ PHASTER. https://phaster.ca/submissions/AEAN00000000.1
- ↑ PHASTER. https://phaster.ca/submissions/AEAN00000000.1
- ↑ Baltrus, D. A., Clark, M., Smith, C. & Hockett, K. L. Localized recombination drives diversification of killing spectra for phage-derived syringacins. ISME Journal 13, 237–249 (2019).