Intermediate

Part:BBa_K2036007

Designed by: Zhangyu Cheng   Group: iGEM16_HUST-China   (2016-09-19)


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]


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Categories
Parameters
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