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	The results showed that there was a significant difference between the group containing the element and the control group, which proved that the element could perform the expected function, that is, tetR could normally express the repressor protein to inhibit the intensity of deGFP.		The results showed that there was a significant difference between the group containing the element and the control group, which proved that the element could perform the expected function, that is, tetR could normally express the repressor protein to inhibit the intensity of deGFP.

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Revision as of 17:32, 17 October 2021

P28-tetO-deGFP

Under the P28 promoter, the Tet operator tetO is expressed along with the production of green fluorescent protein.

Usage and Biology

1.This plasmid has no label ssrA . We use it to investigate the effect of degradation label ssrA on deGFP fluorescent protein.
2.Provide tetO to binding repressor protein and inhibit the production of fluorescent protein.
3.It can be used in other experiments which hope to express fluorescence.

Characterization

This part was mainly combined with tetR to verify its success in suppressing the expression of deGFP. By electroporation of BW25113 and plasmid containing P70-σ28-P28-tetR as well as plasmid containing P28-tetO-deGFP, it was verified that tetR could inhibit the expression of deGFP successfully.

The results showed that there was a significant difference between the group containing the element and the control group, which proved that the element could perform the expected function, that is, tetR could normally express the repressor protein to inhibit the intensity of deGFP.

Design Page

Some research shows that the novel Ptar–tetO hybrid promoter is repressed by tetR,Schematic of an incoherent type-1 feed-forward loop. Transcription unit 1 (TU1) constitutively expresses σ28, which activates the expression of both transcription unit 2 (TU2) and transcription unit 3 (TU3). TU2 produces the tetR repressor protein, which represses the production of GFP from TU3. The simultaneous activation of TU2 and TU3 results in a pulse of GFP when the protease ClpXP is present to degrade the GFP signal.

References

Melissa K. Takahashi et al. Characterizing and prototyping genetic networks with cell-free transcription–translation reactions[J]. Methods, 2015, 86 : 60-72.

Sequence and Features