Difference between revisions of "Part:BBa K862000"
 
(12 intermediate revisions by 2 users not shown)
Line 3: Line 3:
  
 
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).
−
 
  
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
−
 
+
<br/>
 
<!-- -->
 
<!-- -->
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
Line 18: Line 17:
 
<partinfo>BBa_K862000 parameters</partinfo>
 
<partinfo>BBa_K862000 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
 +
 +
===Characterisation===
 +
This part was characterized during the 2012 HS competition by the Heidelberg_LSL team
 +
<br/>
 +
<html>
 +
<center><img src="https://static.igem.org/mediawiki/2012hs/8/80/Bildschirmfoto_2012-06-11_um_23.47.05.png" width="500"/></center>
 +
<p>
 +
<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/>
 +
<br/>
 +
</p>
 +
<br/>
 +
<center><img src="https://static.igem.org/mediawiki/2012hs/c/c7/XGAL_Measurement.PNG" width="635"/></center>
 +
<br/><p>
 +
<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/>
 +
<br/>
 +
<center><img src="https://static.igem.org/mediawiki/2012hs/6/6f/GFP_Measurement.PNG" width="500"/></center>
 +
<p>
 +
<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/>
 +
<br/>
 +
 +
</html>

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


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE 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.