Difference between revisions of "Part:BBa K2036000"
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<partinfo>BBa_K2036000 parameters</partinfo> | <partinfo>BBa_K2036000 parameters</partinfo> | ||
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+ | <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> | ||
+ | [[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 increasedover time and it showed significant difference from CK. | ||
+ | ]] | ||
+ | [[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. | ||
+ | ]] |
Revision as of 15:08, 19 October 2016
CII bacteriophage lambda trascriptional activator
CII is an important part of the bacteriaphage lambda operon. It promotes lysogeny through activation of three phage promoters p(E), p(I) and p(aQ), recognizing a direct repeat sequence TTGCN6TTGC at each. It is an unstable protein in vivo, being rapidly degraded by the host protease HflB (FtsH). This instability is essential for the function of CII in the lysis-lysogeny switch. From NMR and limited proteolysis we show that about 15 C-terminal residues of CII are highly flexible, and may act as a target for proteolysis in vivo. From in vitro transcription, isothermal calorimetry and gel chromatography of CII (1-97) and its truncated fragments CIIA (4-81/82) and CIIB (4-69), we find that residues 70-81/82 are essential for (a) tetramer formation, (b) operator binding and (c) transcription activation. Presumably, tetramerization is necessary for the latter functions.
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]
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-GFPLVAssrAtag as CK to see if CII efficiently activate pRE.