Team:SDU-Denmark/project-r

From 2010.igem.org

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=== Effect ===
=== Effect ===
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Our results from the experiments with semi-solid agar confirm that the biobrick does indeed couple itself to the bacterial chemotaxis pathway and modify the bacterial motility pattern by modifying the tumbling frequency. For further information on this conclusion see [http://2010.igem.org/Team:SDU-Denmark/project-p#Growth_of_bacterial_culture_on_semi-solid_agar_plates characterization of K343007]<br>
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Our results from the experiments with semi-solid agar confirm that the biobrick does indeed couple itself to the bacterial chemotaxis pathway and modify the bacterial motility pattern by modifying the tumbling frequency. For further information on this conclusion see [http://2010.igem.org/Team:SDU-Denmark/project-p#Growth_of_bacterial_culture_on_semi-solid_agar_plates growth of bacteria on semi-solid agar plates]<br>
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The videomicroscopy indicates that blue light with a wavelength around 500nm leads to CheA's autophosphorylation being downregulated. This means that the bacterial tumbling frequency gets reduced and the bacteria will spend an increased time in the "run" mode of propulsion. This means that the bacteria will travel further in a certain amount of time, compared to wildtype bacteria. More details can be found here [http://2010.igem.org/Team:SDU-Denmark/project-p#Videomicroscopy_and_computer_analysis_of_bacterial_motility characterization of K343007]<br>
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The videomicroscopy indicates that blue light with a wavelength around 500nm leads to CheA's autophosphorylation being downregulated. This means that the bacterial tumbling frequency gets reduced and the bacteria will spend an increased time in the "run" mode of propulsion. This means that the bacteria will travel further in a certain amount of time, compared to wildtype bacteria. More details can be found here [http://2010.igem.org/Team:SDU-Denmark/project-p#Videomicroscopy_and_computer_analysis_of_bacterial_motility videomicroscopy]<br>
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The stability of pSB1C3-K343007 is most likely <20 generations, however the stability of pSB3T5-K343007 was not determined (see [http://2010.igem.org/Team:SDU-Denmark/project-p#Stability_assay characterization of K343007])<br><br>
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The stability of pSB1C3-K343007 is most likely <20 generations, however the stability of pSB3T5-K343007 was not determined (see [http://2010.igem.org/Team:SDU-Denmark/project-p#Stability_assay stability assay])<br><br>
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Plasmids expressing K343007 does not seem to hinder the bacterial growth in any way (see [http://2010.igem.org/Team:SDU-Denmark/project-p#Growth_assay characterization of K343007])<br>
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Plasmids expressing K343007 does not seem to hinder the bacterial growth in any way (see [http://2010.igem.org/Team:SDU-Denmark/project-p#Growth_assay growth assay])<br><br>
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== Retinal ==
 
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=== Biobrick assembly and sequence ===
 
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We succesfully assembled the biobrick [http://partsregistry.org/Part:BBa_K343006 K343006], which consists of the TetR repressible promoter R0040 followed by the ribosome binding site B0034, the coding sequence K343002 and the terminators B0010 and B0012.<br>
 
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This was achieved through "standard" silver assembly and confirmed via sequencing with biobrick verification forward 2 (VF2) and verification reverse (VR) primers.<br>
 
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Sequencing results (as .ab1 files):<br>
 
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[http://2010.igem.org/wiki/images/a/ab/Sequencing_K343006.ab1| Sequencing results for K343006]
 
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When comparing the theoretical sequence with the sequence done on the part K343006 the two sequences was identical.
 
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=== Effect ===
 
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The growth assay of ''E. coli'' strain MG1655 transformed with plasmids containing the NinaB brick shows no significant difference from the wild type ''E. coli'' strain MG1655, we did however remove and outlier from the [http://2010.igem.org/Team:SDU-Denmark/project-p#3._Growth_measurement: graph]. [http://2010.igem.org/Team:SDU-Denmark/raw raw data] <br>
 
==Flagella==
==Flagella==
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The growth assay showed no significant difference between the wild type and the cells containing either of the two plasmids.<br>
The growth assay showed no significant difference between the wild type and the cells containing either of the two plasmids.<br>
For further information, raw data and background of the assay see [http://2010.igem.org/Team:SDU-Denmark/project-p#K343004 characterization of K343004] <br><br>
For further information, raw data and background of the assay see [http://2010.igem.org/Team:SDU-Denmark/project-p#K343004 characterization of K343004] <br><br>
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== Retinal ==
 +
=== Biobrick assembly and sequence ===
 +
We succesfully assembled the biobrick [http://partsregistry.org/Part:BBa_K343006 K343006], which consists of the TetR repressible promoter R0040 followed by the ribosome binding site B0034, the coding sequence K343002 and the terminators B0010 and B0012.<br>
 +
This was achieved through "standard" silver assembly and confirmed via sequencing with biobrick verification forward 2 (VF2) and verification reverse (VR) primers.<br>
 +
Sequencing results (as .ab1 files):<br>
 +
<br>
 +
[http://2010.igem.org/wiki/images/a/ab/Sequencing_K343006.ab1| Sequencing results for K343006]
 +
<br><br>
 +
When comparing the theoretical sequence with the sequence done on the part K343006 the two sequences was identical.
 +
 +
=== Effect ===
 +
The growth assay of ''E. coli'' strain MG1655 transformed with plasmids containing the NinaB brick shows no significant difference from the wild type ''E. coli'' strain MG1655, we did however remove and outlier from the [http://2010.igem.org/Team:SDU-Denmark/project-p#3._Growth_measurement: graph]. [http://2010.igem.org/Team:SDU-Denmark/raw raw data] <br>

Revision as of 12:21, 26 October 2010