Difference between revisions of "Part:BBa K2036012"
 
(7 intermediate revisions by 3 users not shown)
Line 3: Line 3:
 
<partinfo>BBa_K2036012 short</partinfo>
 
<partinfo>BBa_K2036012 short</partinfo>
 
<br>
 
<br>
−
CII  ([https://parts.igem.org/Part:BBa_K2036000 BBa_K2036000] ) is a regulatory gene derived from bacteriaphage lambda. It is an activate pRE's expression. And it can be normally degrade by Ftsh in E.coli.
+
CII  ([https://parts.igem.org/Part:BBa_K2036000 BBa_K2036000] ) is a regulatory gene derived from bacteriaphage lambda. It is an activate pRE's expression. And it can be normally degrade by Ftsh in E.coli. This part is designed as CII expression frame with RBS ahead and terminator behind.
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
Line 17: Line 17:
 
<partinfo>BBa_K2036012 parameters</partinfo>
 
<partinfo>BBa_K2036012 parameters</partinfo>
 
<!-- -->
 
<!-- -->
 +
<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 test group and pRE-RBS-GFP-LVAssrAtag 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.]]
 +
 +
 +
 +
<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 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 Flourescence measurement curve above, we can see clearly that GFP level of CIII test circuit increased over time and it showed significant difference from two control groups. It indicates that tandemly expressed CIII can efficiently protect CII from being degraded by Ftsh. ]]

Latest revision as of 06:59, 25 October 2016


RBS-CII-TT
CII (BBa_K2036000 ) is a regulatory gene derived from bacteriaphage lambda. It is an activate pRE's expression. And it can be normally degrade by Ftsh in E.coli. This part is designed as CII expression frame with RBS ahead and terminator behind.

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]


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 test group and pRE-RBS-GFP-LVAssrAtag as CK to see if CII efficiently activate pRE.


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.


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.


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 to test whether CIII can protect CII from being degraded by Ftsh by competitive inhibition.


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