Can I still use pGEFP-C1 vector? - (Aug/17/2006 )
I designed primers to insert the fragment into pEFG-N1. Now I want to insert the fragment into pEGFP-C1, that means, it will express GFP +N-protein-C, because I put ATG into 5’ primer, then there will be an initial codon ATG between GFP and my interest protein. Can I still get the product” GFP+interest protein”?
You can use pEGFP-C1 vector for your purpose . In this case, your insert is expreesed as a fusion protein with GFP taking the ATG of GFP tag. Moreover, pEGFP-C1 vector has stop codon in all 3 reading frames after MCS. So, go ahead.
if u plan to PCR amplify a fragment to insert into pEGFP-C1, make sure the sites match as u had initially planned to insert into pEFG-N1.