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e coli vector and host selection


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#1 mbsea

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Posted 13 April 2012 - 10:53 PM

Hi everybody, i am confused about an issue. I want to express a plant beta glucosidase gene. My gene of interest seems hasno disulfide bonds and soluble at a rate of 58%. And seems no transmembrane protein. I am trying to clone it into pet21a, but when serch some articles, they offer pet32 series for insoluble proteins. Do you think the 58% rate of solubility means i must try a vector which is more effective for insoluble proteins? Origami strains are offered for insoluble proteins but my protein has no disulfide bonds but in pet manual, they say: origami strains help to occur disulfide bonds. Could you please offer me a vector and a host for my experiment because i am very confused now.
Best regards.

#2 hRNA

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Posted 03 May 2012 - 09:20 PM

Hi
First of all, how did you came to know about the percent solubility of ur protein?
Secondly, if it is 58% soluble then any of TOPO expression vectors which provide tags (eg trx tag) can be used for the cloning. the main advantage of TOPO is, it is easy, time saving and no need of restriction digestion.you can express it origami/rosetta strains of E. Coli.




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