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Fusion protein expression

Fusion protein expression - Recombinant Protein Forum

Fusion protein expression - Recombinant Protein Forum. Discuss purification, expression, and isolation of recombinant and cloned proteins here.


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Old 03-09-2011, 09:51 AM
Pipette Filler
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Default Fusion protein expression



Hi,
i fused 3.3 kb fusion gene in pET 28 (C),transformed in BL21 (DE3) pLysS, induced with IPTG 1 mM,4 hrs, did western blot (first gene antibody) ,only my first gene (62 KDa) expressed not the second gene (60 KDa). I should get the blot at 120 KDa size. Both of my protein is un unsoluble form.What may be the reason?
Please suggest ....
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Old 10-18-2012, 09:45 PM
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Default Re: Fusion protein expression

Only about 30% of recombinant proteins expressed in e.coli are soluble from the get go. Alternatively you could purify the insoluble portion and refold the protein into it's native state.
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