Expression system for hydrophobic proteins

Jongrak Kittiworakarn g3937506 at STUDENT2.MAHIDOL.AC.TH
Wed Aug 12 23:57:04 EST 1998


> Hi Helen:

I used GST-fusion to express my hydrophobic peptide. The peptide was in insoluble
form.  I solubilized the inclusion with 4M GdnCl, then I refolded it by dialysis
against 6M 2M, 1M, 0.5M, 0.25M, 0.1M and 0M urea respectively at 4 C (it take 2-6
hr. for each step).For an alternative choice, you may try some E.coli host form
Prof. John Walker (at MRC). Here is his publication about the E.coli :

Miroux B, and Walker JM. Over-production of proteins in Escherichia
coli: mutant hosts that allow synthesis of some membrane proteins and
globular proteins at high levels. J Mol Biol. 1996 ; 260: 289-298.

Please tell me if you get any result by trying these.
Good luck,

Jongrak

> Expression system for hydrophobic proteins
>
> Helen Elina (helina at bio1.lan.mcgill.ca)
> Wed, 12 Aug 1998 16:10:18 -0400
>
>    * Messages sorted by: [ date ][ thread ][ subject ][ author ]
>    * Next message: Frank O. Fackelmayer: "Re: HELP!! transformation problems"
>    * Previous message: rao7 at earthlink.net: "RE: Looking for work in USA!"
>
>   ------------------------------------------------------------------------
>
> To: methods at net.bio.net
>
> From: Helen Elina <helina at bio1.lan.mcgill.ca>
>
> Subject: Expression system for hydrophobic proteins
>
> Date: Wed, 12 Aug 1998 16:10:18 -0400
>
> I am trying to express a hydrophobic protein in E. coli, but have not
> succeeded in doing so. I have tried a GST-fusion but the protein is not
> recovered in the soluble fraction.
> Have any of you had success with other expression systems? Please
> advise me on this matter.
> Thanks!
>
> Helen
>   ------------------------------------------------------------------------
>
>    * Next message: Frank O. Fackelmayer: "Re: HELP!! transformation problems"
>    * Previous message: rao7 at earthlink.net: "RE: Looking for work in USA!"






More information about the Methods mailing list