Difference between revisions of "Part:BBa K862000"
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This is a part for the precise quantification of UV-radiation or radioactive radiation in E. coli (recA+) strains, i.e. BL21(DE3). It consits of a recA Promoter (part BBa_J22106) fused to a LacZ reporter cloned in front of a double terminator (part BBa_K173004). | This is a part for the precise quantification of UV-radiation or radioactive radiation in E. coli (recA+) strains, i.e. BL21(DE3). It consits of a recA Promoter (part BBa_J22106) fused to a LacZ reporter cloned in front of a double terminator (part BBa_K173004). | ||
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| + | <!-- Add more about the biology of this part here | ||
| + | ===Usage and Biology=== | ||
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| + | <!-- --> | ||
| + | <span class='h3bb'>Sequence and Features</span> | ||
| + | <partinfo>BBa_K862000 SequenceAndFeatures</partinfo> | ||
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| + | <!-- Uncomment this to enable Functional Parameter display | ||
| + | ===Functional Parameters=== | ||
| + | <partinfo>BBa_K862000 parameters</partinfo> | ||
| + | <!-- --> | ||
===Characterisation=== | ===Characterisation=== | ||
| − | This part was characterized during the 2012 HS competition by the | + | This part was characterized during the 2012 HS competition by the Heidelberg_LSL team |
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<html> | <html> | ||
<center><img src="https://static.igem.org/mediawiki/2012hs/8/80/Bildschirmfoto_2012-06-11_um_23.47.05.png" width="500"/></center> | <center><img src="https://static.igem.org/mediawiki/2012hs/8/80/Bildschirmfoto_2012-06-11_um_23.47.05.png" width="500"/></center> | ||
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| − | <b>Fig. 1: ONPG assay of Bl21(DE3) irradiated for different times. </b>Both constructs show a strong correlation between the UV-irradiation time and the LacZ activity (production of o-nitrophenol). <br/> | + | <b>Fig. 1: Comparison of parts <a href="https://parts.igem.org/Part:BBa_K862000">BBa_K862000 (precA_LacZ)</a> and <a href="https://parts.igem.org/Part:BBa_K862001">BBa_K862001 (psulA_LacZ)</a> via ONPG assay of Bl21(DE3) UV-irradiated for different times. </b>Both constructs show a strong correlation between the UV-irradiation time and the LacZ activity (as measured by the production of o-nitrophenol). <br/> |
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<center><img src="https://static.igem.org/mediawiki/2012hs/c/c7/XGAL_Measurement.PNG" width="635"/></center> | <center><img src="https://static.igem.org/mediawiki/2012hs/c/c7/XGAL_Measurement.PNG" width="635"/></center> | ||
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| − | <b>Fig. 2: X-gal assay of Bl21(DE3) transformed with | + | <b>Fig. 2: X-gal assay of Bl21(DE3) transformed with <a href="https://parts.igem.org/Part:BBa_K862000">BBa_K862000 (precA_LacZ)</a>, <a href="https://parts.igem.org/Part:BBa_K862001">BBa_K862001 (psulA_LacZ)</a> and <a href="https://parts.igem.org/Part:BBa_K862002">BBa_K862002 (precB_LacZ)</a> and UV-irradiated for different times. </b>All constructs show a strong positive correlation between UV induction time and coloring of the wells. PrecA-LacZ (BBa_K862000) gives the lowest reporter background expression whereas psulA-LacZ (BBa_K862001) gives the highest overall coloring of the samples.<br/> |
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<center><img src="https://static.igem.org/mediawiki/2012hs/6/6f/GFP_Measurement.PNG" width="500"/></center> | <center><img src="https://static.igem.org/mediawiki/2012hs/6/6f/GFP_Measurement.PNG" width="500"/></center> | ||
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| − | <b>Fig. 3: Test of precA-GFP reporter construct by fluorescence microscopy. </b><i>E.coli</i> transformed with our precA-GFP reporter were either UV-irradiated for 30 min or left uninduced. measurement of GFP expression show a 10-fold (!) increase in GFP expression due to UV-irradiation compared to the control. <br/> | + | <b>Fig. 3: Test of precA-GFP reporter construct (part <a href="https://parts.igem.org/Part:BBa_J22106">BBa_J22106</a> cloned into GFP backbone <a href="https://parts.igem.org/Part:BBa_E0040">BBa_E0040</a>) by fluorescence microscopy. </b><i>E.coli</i> transformed with our precA-GFP reporter were either UV-irradiated for 30 min or left uninduced. measurement of GFP expression show a 10-fold (!) increase in GFP expression due to UV-irradiation compared to the control. <br/> |
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Latest revision as of 12:36, 25 June 2012
precA-LacZ-doubleTerminator
This is a part for the precise quantification of UV-radiation or radioactive radiation in E. coli (recA+) strains, i.e. BL21(DE3). It consits of a recA Promoter (part BBa_J22106) fused to a LacZ reporter cloned in front of a double terminator (part BBa_K173004).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterisation
This part was characterized during the 2012 HS competition by the Heidelberg_LSL team
Fig. 1: Comparison of parts BBa_K862000 (precA_LacZ) and BBa_K862001 (psulA_LacZ) via ONPG assay of Bl21(DE3) UV-irradiated for different times. Both constructs show a strong correlation between the UV-irradiation time and the LacZ activity (as measured by the production of o-nitrophenol).
Fig. 2: X-gal assay of Bl21(DE3) transformed with BBa_K862000 (precA_LacZ), BBa_K862001 (psulA_LacZ) and BBa_K862002 (precB_LacZ) and UV-irradiated for different times. All constructs show a strong positive correlation between UV induction time and coloring of the wells. PrecA-LacZ (BBa_K862000) gives the lowest reporter background expression whereas psulA-LacZ (BBa_K862001) gives the highest overall coloring of the samples.
Fig. 3: Test of precA-GFP reporter construct (part BBa_J22106 cloned into GFP backbone BBa_E0040) by fluorescence microscopy. E.coli transformed with our precA-GFP reporter were either UV-irradiated for 30 min or left uninduced. measurement of GFP expression show a 10-fold (!) increase in GFP expression due to UV-irradiation compared to the control.