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I am going to prepare a c-myc fusion protein with the following configuration:

(28-residue signal sequence)-(c-myc)-(GGSGGGSG Linker)-(Protein of Interest (POI))

POI is a transmembrane protein and the 28-residue signal sequence is needed for membrane incorporation in ER.

My concern is the disruption of the c-myc/anti-myc Ab interaction due to the fact that c-myc being fused on both ends. Does anyone have any experience or reference with a similar construct?

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After some further research, I found papers reporting successful co-immunoprecipitation experiments with POI-myc-His fusion proteins.

Moreover, I figured the signal sequence will be cleaved by signal peptidases which will leave the c-myc tag exposed on N-terminus with only an extra residue. I don't think that one extra residue will create a problem.

Here's the tool I used to figure out where the signal sequence will be cleaved: http://www.cbs.dtu.dk/services/SignalP/

Here are the references for POI-myc-His experiments:

  • DOI: 10.1016/j.cub.2006.08.085
  • DOI: 10.1073/pnas.97.2.668
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