Results

Photosensor

Biobrick assembly and sequence

We succesfully assembled the biobrick [http://partsregistry.org/Part:BBa_K343007 K343007], which consists of the TetR repressible promoter R0040 followed by the ribosome binding site B0034, the coding sequence K343003 and the terminators B0010 and B0012.
This was achieved through "standard" silver assembly and confirmed via sequencing with biobrick verification forward 2 (VF2) and verification reverse (VR) primers.
Sequencing results (as .ab1 files):

Sequencing results K343007

The sequence obtained from sequencing is identical with the theoretical sequence of the part, except for a silent mutation in the coding sequence, which does not change the amino acid sequence.

Effect

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 Characterization - K343007 - 1. Growth of bacterial culture on semi-solid agar plates (Experiment 1 and 2)

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 [characterization of K343007]


The stability of pSB1C3-K343007 is most likely <20 generations, however the stability of pSB3T5-K343007 was not determined (see [characterization of K343007])

Plasmids expressing K343007 does not seem to hinder the bacterial growth in any way (see [characterization of K343007])

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.
This was achieved through "standard" silver assembly and confirmed via sequencing with biobrick verification forward 2 (VF2) and verification reverse (VR) primers.
Sequencing results (as .ab1 files):

Sequencing results for K343006

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 graph. raw data

Flagella

Biobrick assembly and sequence

We succesfully assembled the biobrick [http://partsregistry.org/Part:BBa_K343004 K343004], which consists of the TetR repressible promoter R0040 followed by the ribosome binding site B0034, the coding sequence K343000 and the terminators B0010 and B0012.
This was achieved through "standard" silver assembly and confirmed via sequencing with biobrick verification forward 2 (VF2) and verification reverse (VR) primers.
Sequencing results (as .ab1 files):

Sequencing results for K343004

When comparing the theoretical sequence with the sequence done on the part K343004 the two sequences was identical.

Effect

Our results from the motility assay with semi-solid agar confirm that the biobrick does infact increase the motility of the cells. We believe this is caused by hyperflagellation.

The growth assay showed no significant difference between the wild type and the cells containing either of the two plasmids.
For further information, raw data and background of the assay see characterization of K343004

This is where it gets really interesting. Just look what we've made!