Difference between revisions of "Part:BBa K4242012"

Revision as of 02:39, 9 October 2022

Pci+Ppel+RBS+gfp+TT(pHYD-2)

Uses factors Pci(BBa_R0051) and Ppel(BBa_K4242001), strong RBS(BBa_J34801), gfp coding region(BBa_E0040), and TT(BBa_B0015). This part is an easy factor assessment tool. The gfp reporter enables you to find out whether the tandem factor PcI/pel works.

Usage and Biology

During our test of the pHYD-1 (BBa_K4242011), we found that promotor Ppel doesn’t work well in many organisms. To overcome this difficulty, we designed a tandem promoter PcI/pel in which the transcription start site of the constitutive promoter PcI is immediately followed by the FleQ binding site of the promoter Ppel. Transcription of the reporter gene gfp is promoted by cI promoter in the absence of FleQ. We found that the fluorescence of recombinant E. coli/pHYD-2 and S. oneidensis/pHYD-2 was drastically higher than that of the strains harboring pHYD-1 (BBa_K4242011), indicating that the tandem promoter is capable of constitutively promoting gfp expression in the absence of FleQ in both strains.

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
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 874

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 ===Usage and Biology===
-During our test of the pHYD-1 (BBa_K4242011), we found that promotor Ppel doesn’t work well in many organisms. To overcome this difficulty, we designed a tandem promoter PcI/pel in which the transcription start site of the constitutive promoter PcI is immediately followed by the FleQ binding site of the promoter Ppel. Transcription of the reporter gene gfp is promoted by cI promoter in the absence of FleQ. We found that the fluorescence of recombinant <i>E. coli</i>/pHYD-2 and <i>S. oneidensis</i>/pHYD-2 was drastically higher than that of the strains harboring pHYD-1 (BBa_K4242011), indicating that the tandem promoter is capable of constitutively promoting gfp expression in the absence of FleQ in both strains.
+During our test of the pHYD-1 (BBa_K4242011), we found that promotor Ppel doesn’t work well in many organisms. To overcome this difficulty, we designed a tandem promoter PcI/pel in which the transcription start site of the constitutive promoter PcI is immediately followed by the FleQ binding site of the promoter Ppel. Transcription of the reporter gene <i>gfp</i> is promoted by cI promoter in the absence of FleQ. We found that the fluorescence of recombinant <i>E. coli</i>/pHYD-2 and <i>S. oneidensis</i>/pHYD-2 was drastically higher than that of the strains harboring pHYD-1 (BBa_K4242011), indicating that the tandem promoter is capable of constitutively promoting <i>gfp</i> expression in the absence of FleQ in both strains.
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