Difference between revisions of "Part:BBa K2036007"
 
(8 intermediate revisions by 2 users not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2036007 short</partinfo>
 
<partinfo>BBa_K2036007 short</partinfo>
  
−
CII is a transcriptional activator of pRE, but it is normally degrade by Ftsh, and in this case,tandom expression maybe useful to activate pRE.
+
CII is a transcriptional activator of pRE, but it is normally degrade by Ftsh, and in this case,tandem expression is useful to activate pRE.
 +
 
 +
<h2>Protein&promoter</h2>
 +
<p>--CII and pRE</p>
 +
<br>
 +
<p>
 +
CII ([https://parts.igem.org/Part:BBa_K2036000 BBa_K2036000]) functions as a transcriptional activator to direct promoter RE, so we constructed CII-TT-pRE-RBS-GFP-LVAssrAtag as the test group and pRE-RBS-GFP-LVAssrAtag as CK to see whether CII efficiently activates pRE.
 +
</p>
 +
<br>
 +
[[File:T--HUST-China--CII-pRE_plate.png|800px|thumb|center|Fig1: According to the Fluorescence measurement curve above, we can see clearly that GFP level increased over time and it showed significant difference from CK.]]
 +
<br>
 +
[[File:T--HUST-China--Experiments-CII-pRE_Flou-detec.png|800px|thumb|center|Fig2: We also did Fluorescence microscope detection after 30, 120 and 240 minutes' induction. According to the figure above, we can tell qualitatively that pRE leakage are at relative low level and CII can efficiently activate the promoter.]]
 +
<h2>Protein&protein reaction</h2>
 +
 
 +
<p>
 +
We had submitted and documented RBS-CIII-RBS-CIII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag ([https://parts.igem.org/Part:BBa_K2036014 BBa_K2036014]) and RBS-CII-RBS-CII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag ([https://parts.igem.org/Part:BBa_K2036015 BBa_K2036015]).These two parts were constructed to test whether CIII can protect CII from being degraded by Ftsh by competitive inhibition.
 +
</p>
 +
<br>
 +
[[File:T--HUST-China--CIII%26Ftsh.png|800px|thumb|center|Fig3: According to the Fluorescence measurement curve above, we can see clearly that GFP intensity of the CIII test circuit increases over time and it shows significant difference from two control groups. It indicates that tandemly expressed CIII can efficiently protect CII from being degraded by Ftsh. ]]
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 17: Line 35:
 
<partinfo>BBa_K2036007 parameters</partinfo>
 
<partinfo>BBa_K2036007 parameters</partinfo>
 
<!-- -->
 
<!-- -->
−
<h2>Protein&promoter</h2>
 
−
<p>--CII and pRE</p>
 
−
<br>
 
−
<p>
 
−
CII (BBa_K2036000) functions as a transcriptional activator to direct promoter RE, so we constructed CII-TT-pRE-RBS-GFP-LVAssrAtag as test group and pRE-RBS-GFPLVAssrAtag as CK to see if CII efficiently activate pRE.
 
−
</p>
 
−
<br>
 
−
[[File:T--HUST-China--CII-pRE_plate.png|800px|thumb|center|Fig1: According to the Flourescence measurement curve above, we can see clearly that GFP level increased over time and it showed significant difference from CK.]]
 
−
<br>
 
−
[[File:T--HUST-China--Experiments-CII-pRE_Flou-detec.png|800px|thumb|center|Fig2: We also did Fluorescence microscope detection after 30, 120 and 240 minutes induction. According to the figture below, we can tell qualitively that pRE leakage are at relative low level and CII can efficiently activate the promoter.]]
 

Latest revision as of 05:22, 25 October 2016


RBS-CII-RBS-CII, tandem expression

CII is a transcriptional activator of pRE, but it is normally degrade by Ftsh, and in this case,tandem expression is useful to activate pRE.

Protein&promoter

--CII and pRE


CII (BBa_K2036000) functions as a transcriptional activator to direct promoter RE, so we constructed CII-TT-pRE-RBS-GFP-LVAssrAtag as the test group and pRE-RBS-GFP-LVAssrAtag as CK to see whether CII efficiently activates pRE.


Fig1: According to the Fluorescence measurement curve above, we can see clearly that GFP level increased over time and it showed significant difference from CK.


Fig2: We also did Fluorescence microscope detection after 30, 120 and 240 minutes' induction. According to the figure above, we can tell qualitatively that pRE leakage are at relative low level and CII can efficiently activate the promoter.

Protein&protein reaction

We had submitted and documented RBS-CIII-RBS-CIII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag (BBa_K2036014) and RBS-CII-RBS-CII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag (BBa_K2036015).These two parts were constructed to test whether CIII can protect CII from being degraded by Ftsh by competitive inhibition.


Fig3: According to the Fluorescence measurement curve above, we can see clearly that GFP intensity of the CIII test circuit increases over time and it shows significant difference from two control groups. It indicates that tandemly expressed CIII can efficiently protect CII from being degraded by Ftsh.

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]