- Grew up plasmids (pCRII-MTM1, pCRII-MTM1-C375S from Francis; already sequenced)
- Cut prey (pACT2) with BamH1/XhoI, and pCRII-MTM1 with BamH1/SalI
- Cut bait (pFBT9) and pCRII-MTM1 with Sal1 and BamH1 (sequential digestion)
- Cut prey (pACT2) with BamH1/XhoI and pCRII-MTM1-C375S with BamH1/SalI
- Cut bait (pFBT9) and pCRII-MTM1-C375S with BamH1 and Sal1 (sequential digestion)
- Ligation and growing pACT2-MTM1, pACT2-MTM1-C375S, pFBT9-MTM1 and pFBT9-MTM1-C375S.
- Two Hybrid Analyses
- For Co-IP, Co-IF:
Digest:
pCMV-HA with Bgl II and Xho1 and pFBT9-hMTM1 with BamH and Sal1.
pEFGP-C2 with Bgl II and Sal1 and pFBT9-hMTM1 with BamH and Sal1.