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how to make a recombinant enzyme refolded


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#1 joy123

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Posted 15 March 2012 - 01:18 PM

Hi, I am interested in an enzyme in a G(+) bacteria. It natually sits on cell membrane.

I have cloned it in E.coli. But the recombinant protein didn't have enzyme activity (I checked it by loading E coli on zymogram).

My question is: how to make a recombinant enzyme have activity? Does the procedure differs a lot for different enzymes, or there is a universal procedure?

Thanks very much! Look forward to hear from you!

#2 Inmost sun

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Posted 03 April 2012 - 11:47 AM

you may find out an in vitro protocol for reconstitution (f.i. Novogene)

or try coexpression with a chaperon/HSP

#3 joy123

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Posted 04 April 2012 - 09:20 AM

Thanks! I am working on it.




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