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PtasA

Part:BBa_K1701003

Designed by: Wei Wei   Group: iGEM15_Nanjing-China   (2015-09-08)

PtasA

The PtasA gene is the promoter of the TasA gene, and the TasA gene codes for the protein TasA. The TasA protein, in Bacillus subtilis, is one of the major components of the extracellular matrix, which polymerizes into amyloid-like fibers.

We get the sequence of PtasA from the whole genome shotgun sequence of Bacillus subtilis subsp. subtilis str. NCIB 3610 chromosome.

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
    COMPATIBLE 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.

Ptasa1.png

Ptasa02.png

Ptasa03.png

Figure 1. Three pathways designed by our group involving PtasA part.

We could also examine the ability of the expression by using EGFP, the fluorescence protein, and the pathway is shown in Figure 2.

Ptasa2.png

Figure 2. The pathway used for fluorescence measurement.

Results

B. subtilis cells carrying fusion proteins (TasA-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 (TasA-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.

Ptasa3.png

Ptasa4.png

Figure 3. Fluorescence measurement of B. subtilis cells containing the fusion proteins. a) Fluorescence intensity of the fusion proteins Pveg-TasA-EGFP and Ptas-TasA-EGFP. c) A visible photograph of a).


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Categories
//chassis/prokaryote/bsubtilis
Parameters
biologyBacillus subtilis