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Part:BBa_R0051:Experience
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how you used this part and how it worked out.
Applications of BBa_R0051
User Reviews
UNIQ1a4f15695f55f5f3-partinfo-00000000-QINU
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Antiquity |
This review comes from the old result system and indicates that this part worked in some test. |
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Tokyo_Tech |
Transformation and o/n culture is Okay, but we couldn't extract plasmid by miniprep. We couldn't detect band by Electrophoresis. We don't know why, but this occured 2 times. |
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Aberdeen_Scotland 2009 |
The initial gel worked although the obvious confirmation was not possible due to the very small fragment size (49bp).But later it was confirmed following its usage by forming the part BBa_K182002 through the PCR gel analysis and the sequencing.The plasmid miniprep also worked. |
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Seu_O_China 2012 |
BBa_R0051 does not work well due to its rather small size. We transformed it for three times, and none of them succeed. We designed primers to PCR it out from K091230 successfully, but failed to perform the digestion since it is too small to extract from gel. It may work better if adding some nonsense sequence before the promoter.
Device DesignIn this design, we used promoter PcstA and an intermediate regulator cI, as PcstA is activated by CRP in low glucose concentration, whereas we need a device boosted by high concentration of glucose. As you can see, in this pathway, protein cI will be expressed when CRP is activated under low glucose concentration.The large amount of cI repressed the promoter R0051 heavily. On the contrary, when glucose concentration is high, the promoter is derepressed.
In this figure, the fluorescence of RFP generator promoted by PcstA decreased when concentration of glucose rose, meanwhile, the fluorescence of the indirect regulatory device increased with the glucose concentration, indicating that the device works as expected.In another word, promoter R0051 could be repressed by protein cI of lambda phage
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