How can the target gene be transferred from the pCYB vectors into the pTYB vectors?
If the gene of interest has previously been inserted into one of the pCYB vectors, the gene of interest, along with part or all of the intein-CBD coding region, can be transferred into the C-terminal fusion pTYB vectors (pTYB 1,2,3 and 4) by using the compatible unique restriction sites that exist between the two vectors (See table below). Be sure that the sites used are not present in the gene of interest. Furthermore, pMYB5, a control plasmid may be used for cloning a gene of interest. Replacing the malE gene sequence (1.1Kb) between the NdeI and XhoI sites by an open reading frame leaves three extra amino acid residues (LeuGluGly) at the C-terminus of the target protein following cleavage.
