- Subcloned hMTMR3 into bait and prey:
- Cut prey (pACT2) and pAcG2T-K2 with EcoRI. Check orientation
- Cut bait (pFBT9) and pGEMT-Kiaa0371 with EcoR1 and SaI1 (sequencial)
- pACT2-KC413S. Not Done (different frame)
- Cut bait (pFBT9) and pGEMT-KC413S with EcoR1 and Sal1 (sequencial)
- Ligation and transform into electro-DH5 and grow pACT2-hMTMR3, pFBT9-hMTMR3
and pFBT9-hMTMR3-C413S.
- Two Hybrid System
- For Co-IP, Co-IF:
We’ve got already pEGFP-C2-hMTMR3, pEGFP-C413s, HA-hMTMR3 and HA-C413S.
They were subcloned from pGEMT-Kiaa0371 and its mutant pGEMT-C413S anfter sequencing