Also worth trying a lower IPTG level ~ 0.1mM and or different media,
e.g. TB, SOC or m9 instead of LB, it's all a bit random but sometimes
these things can make a difference. Adding 3% EtOH on induction
worked for me once, it's supposed to shock the cells and stimulate
chaperone production, I was surprised I must say. Funny old world......
Giles Robertson D.Phil.
[log in to unmask]
Tel: 02076316813
School of Crystallography,
Birkbeck College,
University of London,
Malet Street,
London, WC1E 7HX.
On 2 Apr 2008, at 12:19, Brenda Patterson wrote:
> Lower temperature, use chaperones (e.g. TAKARA set), refolding?
>
>
> Quoting shivesh kumar <[log in to unmask]>:
>
>> Dear all,
>> Sorry for the off-topic question...
>> What can be done to avoid a protein going inside inclusion
>> body.The gene is
>> cloned in pET30a with C-ter his tag and expressed in BL21-DE3
>> from 37 to
>> 18C for 3-4 hr with .5mM of IPTG,it is going to inclusion body.All
>> suggestions are welcome.
>> Thanx in advance.
>> Shivesh
>>
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