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Need help to find a protein expression protocol for cysteine crosslinked dimers

Need help to find a protein expression protocol for cysteine crosslinked dimers - Protocols and Methods Forum

Need help to find a protein expression protocol for cysteine crosslinked dimers - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 03-27-2008, 09:16 PM
Nirosha Gunasekara
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Default Need help to find a protein expression protocol for cysteine crosslinked dimers



Hello All,

I need help to choose a vector to express a cysteine cross linked
dimeric fragment and the monomeric fragment of a protien for NMR
structure determination. Since I need to express just a part of a
protein what are the limitations (or things to consider) when choosing
an expression vector as well as purification of dimeric and monomeric
proteins? Which vector(s) may be good candidates? Does choosing a
vector depend on the cysteine cross linking procedure? I am wondering
whether vector, pEZZ18 (Amersham Bio) can be used to express just a
part of a protien as I read that the expression by pEZZ18 is
controlled by the lacUV5 and protein A promoters and is not inducible.
I am new to molecular cloning and could you please clarify this as
well. If you could suggest a common protocol for the expression of
cysteine cross linked dimers or provide me with a direction to look
for that would be ideal. All your help is greatly appreciated.

Thank you very much!
Nirosha


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