Part:BBa_K4202003
This part is a fusion protein of mRFP and a engineered protein named SpyCatcher
SpyCatcher-mRFP is a self-designed fusion protein. mRFP is a engineered fluorescent protein. Thus, it`s N and C terminus are allowed to link to other proteins. So we connect SpyCatcher to theN terminus of mRFP through GS sequence to construct SpyCatcher-mRFP. In addition, the N-terminal fusion protein with 6*His tag can be used for protein purification and other experiments.
In our experiment, we hope to use this part and our another part (BBa_K4202004) to verify the feasibility of SpyCatcher-SpyTag system by protein experiments. The protein was also used for experimental validation of the biological scaffold module of our experiment in which we used fluorescence microscopy to characterize the module.
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]
Result:
The plasmid PET-28a(+)-SpyCatcher-mRFP was synthesized by Genscript. And the plasmid wai transferred into BL21(DE3).We set several experience to explore the proper conditions for protein expression. And we finally choose the 25 oC, 220rpm ,0.1mM IPTG, to induce protein.
Then we purified this protein by His-tag Protein Purification Kit.
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