In the case of C-terminal fusion vectors, if the EcoRI site is used as the 3´ cloning site, which vector should be chosen?
The residues (Ser Ser) encoded by the polylinker in pTXB1 or pTYB1 and pTXB3 or pTYB3 preceding the intein results in poor cleavage. Please note that the SapI site can be used to introduce residues favorable for cleavage. If a 3´ cloning site other than the SapI site is chosen, either the pTYB2 or pTYB4 vectors should be used since glycine is the native residue at the N-terminal splice junction of the yeast intein (Sce VMA), thus optimizing cleavage reaction.
Related Questions
- In the case of the C-terminal fusion vectors (pTYB1,2,3 and 4), which residues at the C-terminal of the target protein may inhibit cleavage or cause in vivo cleavage?
- In the case of C-terminal fusion vectors, if the EcoRI site is used as the 3´ cloning site, which vector should be chosen?
- How to design PCR Primers for cloning into C-terminal fusion vectors pTXB, pTYB (pTYB 1,2,3 and 4), or pCYB vectors?
