Part:BBa_K1701002
PcotC
The PcotC gene is the promoter of the CotC gene, and the CotC gene codes for the protein CotC.The CotC protein, in Bacillus subtilis, is one of the the protein component of its spore coat, such as Cot B, Cot C, Cot F, and can be used as a fusion partner to express proteins or polypeptides on the spore surface.
We get the sequence of PcotC from the whole genome shotgun sequence of Bacillus subtilis subsp. subtilis str. NCIB 3610 chromosome.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
It is a weak promoter. We use this promoter to initiate the expression of the following pathways in Figure 1.
Figure 1. Three pathways designed by our group involving PcotC part.
We could also examine the ability of the expression by using EGFP, the fluorescence protein, and the pathway is shown in Figure 2.
Figure 2. The pathway used for fluorescence measurement.
Results
B. subtilis cells carrying fusion proteins (CotC-EGFP) were fixed for visualization under a fluorescence microscope. The same batch of B. subtilis cells without fusion proteins was used as s control. Strong fluorescence was observed when the B. subtilis cells expressed the fusion proteins (CotC-EGFP), whereas no fluorescence was detected for the control group. Taken together, these results confirmed the fusion proteins can express well on the surface of B. subtilis. The following pictures show the results of fluorescence measurements.
Figure 3. Fluorescence measurement of B. subtilis cells containing the fusion proteins. b) Fluorescence intensity of the fusion proteins Pveg-CotC-EGFP and Pcot-CotC-EGFP. c) A visible photograph of b).
| biology | Bacillus subtilis |