Part:BBa_K2830016:Experience

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Applications of BBa_K2830016

To increase the amount of cjcas9 protein in cytoplasm transfers to the periplasm, therefore, increase the chance of the protein to be warped by OMV, torA signal peptide is added in front of the coding sequence of cjcas9.

To test the function of torA, GFP-mut3 was used to instead of cjcas9, since we can observe the effect of torA by measuring the fluorescence in cytoplasm and periplasm respectively to determine the amount of GFP-mut3 protein transferred.

Osmoshock was done to separate the cytoplasm and periplasm of GFP-mut3 protein carried bacteria culture. The result of testing fluorescent value in Figure 1.1 and 1.2 shows that the torA signal peptide significantly increases the efficiency of transfer GFP-mut3 into the periplasm in both kinds of bacteria.

https://parts.igem.org/File:Figure1-1.png https://parts.igem.org/File:Figure1-2.png

Figure1: Fluorescent value in periplasm / total fluorescent value (PF/TF) The fluorescent value in cytoplasm and periplasm is measured respectively and the total value is calculated by adding them together. To compare the results, the ratio between PF and TF is calculated and shown in the diagrams. Figure1.1 and Figure1.2 are experiments done with same method.

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