Team:Imperial College/Biobricks

From 2008.igem.org

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{{Imperial/Box2|Biobrick Characterisation in ''B.subtilis''|
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{{Imperial/Box2|Biobrick Characterisation in ''B. subtilis''|
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This year Imperial College has added 45 new ''B.subtilis'' specific parts to the registry. In addition we sort to characterise some of these parts, including the constitutive promoter and ribosome binding site '''pveg-spoVG'''. The testing constructs we developed for this are shown below:
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The testing constructs we developed for characterisation of one of our promoter-RBS combinations are shown below:
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[[Image:Biobricks Imperial.PNG|center|500px]]
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{{Imperial/Box1|Summary of Data|
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The promoter and RBS combinations were characterised by measuring the steadty-state levels expression of fluorescent proteins in ''B.subtilis''. Cultures of ''B.subtilis'' transformed with the test constructs and non-transformed ''B.subtilis'' (control) were grown to the mid-log phase and measured using a plate reader.
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[[Image:Biobricks Imperial.PNG|center|500px]]
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{{Imperial/Box1|Summary of Data|The promoter and RBS combinations were characterised by measuring the expression of fluorescent proteins in ''B. subtilis''. Cultures of ''B. subtilis'' transformed with the test constructs and non-transformed ''B. subtilis'' (control) were grown to the mid-log phase. Fluorescence and O.D.<sub>600</sub> were both measured using a plate reader to generate fluorescence levels of the various samples. To make this data more generic, the fluorescence data were normalized to cell number using the O.D.<sub>600</sub> and a '''calibration curve of O.D.<sub>600</sub> vs colony forming units''', as explained in the diagram below:
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[[Image:Fluorescence B.subtilis1.PNG]]
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[[Image:Fluorescence B.subtilis.PNG]]
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[[Image:Calibration Picture.PNG|600px|center]]
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This calibration curve allowed us to produce the final data as shown below. The following graph clearly shows that ''B. subtilis'' transformed with GFP and RFP BioBricks are fluorescing at the corresponding excitation and emission wavelengths.
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[[Image:Fluorescence B.subtilis.PNG|400px|center]]
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{{Imperial/EndPage|Major_Results|Major_Results}}

Latest revision as of 02:29, 30 October 2008


Biobrick Characterisation in B. subtilis

The testing constructs we developed for characterisation of one of our promoter-RBS combinations are shown below:

Biobricks Imperial.PNG

Summary of Data

The promoter and RBS combinations were characterised by measuring the expression of fluorescent proteins in B. subtilis. Cultures of B. subtilis transformed with the test constructs and non-transformed B. subtilis (control) were grown to the mid-log phase. Fluorescence and O.D.600 were both measured using a plate reader to generate fluorescence levels of the various samples. To make this data more generic, the fluorescence data were normalized to cell number using the O.D.600 and a calibration curve of O.D.600 vs colony forming units, as explained in the diagram below:

Calibration Picture.PNG


This calibration curve allowed us to produce the final data as shown below. The following graph clearly shows that B. subtilis transformed with GFP and RFP BioBricks are fluorescing at the corresponding excitation and emission wavelengths.

Fluorescence B.subtilis.PNG


Retrieved from "http://2008.igem.org/Team:Imperial_College/Biobricks"