Part:BBa_K2036011
pRE-GFP-LVAssrAtag
It is a GFP generator,and the production of GFP will be activated by a certain level of CII or CII with help of CIII in E.coli.
HUST-China 2016 built this circuit to characterize CII and pRE interaction with test group: RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag (BBa_K2036013) and RBS-CII-RBS-CII-RBS-CII-TT-pRE-RBS-GFP-LVAssrAtag (BBa_K2036015).
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 whether CII can efficiently activate pRE.
Preliminary experiments of LVAssrAtag
In order to prove that our toolkit is efficient to switch two interest genes’ expression from GFP to RFP and to eliminate the accumulation of expressed protein to interfere our measurement. We fused a degradation tag at the amino terminal of our reporter. And we used plac from the Registry(BBa_J04500) to characterize the degradation tag LVAssrA. We use IPTG with final concentration of 1mM to induce the GFP-LVAssrAtag and measure the relative fluorescence through plate reader with excitation wavelength 495nm.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 719
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