2 replies to this topic

[tantao]

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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[rkay447]

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.

[tantao]

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,   Thanks very much!