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help with pCMV-myc vector - (Mar/20/2010 )

Hi all,
I have a question about clontec pCMV-myc vector. I want to clone a gene into EcoRI and NotI sites to form a fusion protein with myc. But I want to know if the sequence ahead of EcoRI site (begin at 868) will affect my protein structure. Thanks a lot!
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-tantao-

It is impossible to say for certain what this linker region (or the myc tag in fact) will do to the native conformation of your protein. Anytime you tag a protein you run the risk of disrupting structure, function, localization, interactions, stability, ect... this is why you need to make sure you are doing all the appropriate controls. I do prefer small tags such as myc or HA to help minimize this risk. The extra sequence between the myc tag and your protein is going to be about 8 amino acids so it is unlikely to disrupt structure but you can always run the tagged protein through structure prediction software to see the likelihood for changes.

-rkay447-

rkay447 on Mar 21 2010, 11:01 AM said:

It is impossible to say for certain what this linker region (or the myc tag in fact) will do to the native conformation of your protein. Anytime you tag a protein you run the risk of disrupting structure, function, localization, interactions, stability, ect... this is why you need to make sure you are doing all the appropriate controls. I do prefer small tags such as myc or HA to help minimize this risk. The extra sequence between the myc tag and your protein is going to be about 8 amino acids so it is unlikely to disrupt structure but you can always run the tagged protein through structure prediction software to see the likelihood for changes.

rKay447, :lol: Thanks very much!

-tantao-