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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”?

-gingerman-

QUOTE (gingerman @ Aug 17 2006, 12:13 PM)
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”?



Hi,
You can use pEGFP-C1 vector for your purpose smile.gif . 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.

masca

-masca-

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.

-scolix-