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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. | 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. | ||
Latest revision as of 17:34, 17 October 2021
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
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
- 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 SapI site found at 487
Illegal SapI.rc site found at 512
Illegal SapI.rc site found at 668