Part:BBa_K322127:Experience

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Applications of BBa_K322127

This part was used in the project of the TU Munich 2011 in two ways:

• as a template for isolation of the red light sensor using PCR. This resulted in the generation of part BBa_K568000, which is the red light sensor without a reporter construct. Therefore, Part BBa_K568000 is an improvement of BBa_K322127, because it improves the ease-of-use. The red light sensor can now be combined with any reporter construct.

• In testing/characterizing part BBa_K568000. To characterize BBa_K568000 and to examine BBa_K322127 a Miller Assay was conducted after transformation of the parts into E. coli CP919. A cph8 part (kindly provided by team Uppsala-Sweden) was used as a negative control. All cells were incubated under three conditions: Irradiation with light of 626 nm ("shut down wavelength"), irradiation with light of 710 nm ("induction wavelength") and in the dark. The measuring was conducted over 210 minutes and then aborted, because no difference between the samples was visible. This means that parts BBa_K322127 and BBa_K568000 did not work as expected in our set-up.

• For further charactisation we used the red light sensor with GFP. We tested its function in JT2. The test of the assay was repeated several times. Our results indicate that the part is independed of the different light conditions active.

This indicates that this part is not sensitive to red light only and not working in the expected manner. For further details see Results in Team TU Munich 2011 wiki.

We suppose this is due to incorrect choice of RBS in the recovery process of this part.

User Reviews

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