Part:BBa_K3885341

P70-σ28-P28-deGFP

In response to the P70 promoter, the transcription factor σ 28 is expressed and activates expression from the P28 promoter

Usage and Biology

To enhance the expression intensity of each component, we use σ28 as the promoter factor. σ28 is a special transcriptional factor concerning the fliA, which is an alternate sigma factor for the class 3 flagella operons, does not exist in Cell-Free system. To verify the functional availability of σ28, we constructed P70a-σ28-P28-deGFP.

Characterization

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This part is used to express and verify the functionality of σ28, so that σ28 will function normally in a Cell-Free system. Through the electroporation experiment, we conducted intra-bacterial verification. The results showed that the fluorescence results of the group containing the part were significantly different from those of the control group (Figure 1 a/b and Figure 2), which proved that the part can perform the expected function, meaning that σ28 can be normal expressed.

Design Page

P28 is a stronger promoter than P70a, and P28 requires σ28 as a promoter factor. There is no σ28 in a Cell-Free system, and we need to express σ28 and verify its functionality, so we constructed P70a-σ28-P28-deGFP part.

References

Larson, M. H., Greenleaf, W. J., Landick, R., and Block, S. M. (2008) Applied force reveals mechanistic and energetic details of transcription termination, Cell 132, 971­982.

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]

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