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EGFP fusion proteins

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Topic Started by galileo_galilei
on 10/2/2010 5:36 AM   
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Hello, I am trying to express a EGFP fusion protein with MY-PROTEIN.  Does anybody have a preference for fusion at  C-terminal or N-terminal?  How about a linker protein between the egfp and MY-PROTEIN?  any suggestions?  many thanks


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Pippuri
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Posted By Pippuri
on 10/3/2010 9:49 AM   
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 Hi galileo_galilei,

Whether your protein prefers a fusion at the C- or N- terminus is largely case-dependent. But here are some questions you can ask yourself before coming up with a conclusion-

Do you know any structural information on this protein? If not, based of the primary sequence, what kind of info you can get? Does it has a specific function found at either the C- or N- terminus that introducing a fusion might kill that specific function? Take an enzyme with a catalytic domain at the C-terminus for instance. Adding a C-terminus fusion might disrupt the catalytic activity which renders the construct useless... If this protein of yours has been reported in the literature, you may find some clues by digging into the literature... If not, you really have to make an educated guess, and test it experimentally...

Hope this helps and good luck! 
Same thing actually apply to the addition of the linker....




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Posted By galileo_galilei
on 10/3/2010 20:25 PM   
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thanks,



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