Re: [ccp4bb] Cys auxotroph

[Paula Salgado]

For those interested in our paper using non-auxotroph strains for SE-Cys
labelling, it's now published:

http://journals.iucr.org/d/issues/2011/01/00/dz5216/index.html

Please cite
this reference if you use our method in the future.

Best regards
Paula

On 20 October 2010 12:00, Paula Salgado  wrote:

> We are still waiting for proofs, so it might be a few weeks before its
> published. If anyone is interested, please email me requesting a preview
> version.
>
> Paula
>
>
> PS: Francois, we did cite and used your work as starting point.
>
> On 20 October 2010 10:39, francois@cbs.cnrs.fr <
> francois@cbs.cnrs.fr> wrote:
>
>> Hi Paula
>>
>> Also our publication: http://www.ncbi.nlm.nih.gov/pubmed/14604526
>> We can provide you the Cys-auxotroph strain of E. coli
>>
>> Francois
>>
>>
>> === Francois Hoh, PhD 
>> CBS Phone : (33) 4 67 41 77 06
>> 29 rue de Navacelles  Fax : (33) 4 67 41 79 13
>> 34090 MONTPELLIER francois@cbs.cnrs.fr
>> France
>> www.cbs.cnrs.fr
>>
>>
>>
>> > I would also try non-auxotrophic strains. We have recently had
>> successful
>> > double SeCys + SeMet using BL21. Paper has just been accepted in Acta D,
>> > should be published soon.
>> >
>> > Paula
>> >
>> > On 18 October 2010 16:05, Yogesh Gupta 
>> wrote:
>> >
>> >> Does anyone know a commercial source of  Cys-auxotroph strain of E.
>> coli
>> >> to
>> >> prepare a doubly (Se-Cys + Se-Met) labeled protein? I would appreciate
>> >> if
>> >> you can share your experience if you have labeled your protein in this
>> >> way
>> >> to improve Se signal for phasing.
>> >>
>> >> Thanks,
>> >> -Yogesh
>> >>
>> >> --
>> >> --
>> >> Yogesh K. Gupta, Ph.D.
>> >> Dept of Structural & Chemical Biology
>> >> Mount Sinai School of Medicine
>> >> Icahn Medical Institute
>> >> 1 Gustave L. Levy Place, Box 1677
>> >> New York, NY, USA 10029
>> >>
>> >> Tel:+1 212-659-8639
>> >> --
>> >>
>> >
>>
>>
>>
>>
>>
>>
>>
>>
>

Re: [ccp4bb] Cys auxotroph

[Paula Salgado]

We are still waiting for proofs, so it might be a few weeks before its
published. If anyone is interested, please email me requesting a preview
version.

Paula


PS: Francois, we did cite and used your work as starting point.

On 20 October 2010 10:39, francois@cbs.cnrs.fr  wrote:

> Hi Paula
>
> Also our publication: http://www.ncbi.nlm.nih.gov/pubmed/14604526
> We can provide you the Cys-auxotroph strain of E. coli
>
> Francois
>
>
> === Francois Hoh, PhD 
> CBS Phone : (33) 4 67 41 77 06
> 29 rue de Navacelles  Fax : (33) 4 67 41 79 13
> 34090 MONTPELLIER francois@cbs.cnrs.fr
> France
> www.cbs.cnrs.fr
>
>
>
> > I would also try non-auxotrophic strains. We have recently had successful
> > double SeCys + SeMet using BL21. Paper has just been accepted in Acta D,
> > should be published soon.
> >
> > Paula
> >
> > On 18 October 2010 16:05, Yogesh Gupta  wrote:
> >
> >> Does anyone know a commercial source of  Cys-auxotroph strain of E. coli
> >> to
> >> prepare a doubly (Se-Cys + Se-Met) labeled protein? I would appreciate
> >> if
> >> you can share your experience if you have labeled your protein in this
> >> way
> >> to improve Se signal for phasing.
> >>
> >> Thanks,
> >> -Yogesh
> >>
> >> --
> >> --
> >> Yogesh K. Gupta, Ph.D.
> >> Dept of Structural & Chemical Biology
> >> Mount Sinai School of Medicine
> >> Icahn Medical Institute
> >> 1 Gustave L. Levy Place, Box 1677
> >> New York, NY, USA 10029
> >>
> >> Tel:+1 212-659-8639
> >> --
> >>
> >
>
>
>
>
>
>
>
>

Re: [ccp4bb] Cys auxotroph

[Paula Salgado]

I would also try non-auxotrophic strains. We have recently had successful
double SeCys + SeMet using BL21. Paper has just been accepted in Acta D,
should be published soon.

Paula

On 18 October 2010 16:05, Yogesh Gupta  wrote:

> Does anyone know a commercial source of  Cys-auxotroph strain of E. coli to
> prepare a doubly (Se-Cys + Se-Met) labeled protein? I would appreciate if
> you can share your experience if you have labeled your protein in this way
> to improve Se signal for phasing.
>
> Thanks,
> -Yogesh
>
> --
> --
> Yogesh K. Gupta, Ph.D.
> Dept of Structural & Chemical Biology
> Mount Sinai School of Medicine
> Icahn Medical Institute
> 1 Gustave L. Levy Place, Box 1677
> New York, NY, USA 10029
>
> Tel:+1 212-659-8639
> --
>

Re: [ccp4bb] Cys auxotroph

[M T]

I think you should look at pubmed id 14604526, it works well for me. You should 
obtain the cells if you contact the authors. 

Michel.

Le 18 oct. 2010 à 17:27, Daniel Bonsor  a écrit :

> Have you looked at the Keio collection. Though they are not compatible with 
> T7 promoters, you can use the λDE3 Lysogenization Kit to add the T7 
> polymerase. 
> 
> A possible source is from Yale http://cgsc.biology.yale.edu/Auxotrophs.php 
> though of course there are others.
> 
> 
> Dan

Re: [ccp4bb] Cys auxotroph

[Daniel Bonsor]

Have you looked at the Keio collection. Though they are not compatible with T7 
promoters, you can use the λDE3 Lysogenization Kit to add the T7 polymerase. 

A possible source is from Yale http://cgsc.biology.yale.edu/Auxotrophs.php 
though of course there are others.


Dan

[ccp4bb] Cys auxotroph

[Yogesh Gupta]

Does anyone know a commercial source of  Cys-auxotroph strain of E. coli to
prepare a doubly (Se-Cys + Se-Met) labeled protein? I would appreciate if
you can share your experience if you have labeled your protein in this way
to improve Se signal for phasing.

Thanks,
-Yogesh

-- 
--
Yogesh K. Gupta, Ph.D.
Dept of Structural & Chemical Biology
Mount Sinai School of Medicine
Icahn Medical Institute
1 Gustave L. Levy Place, Box 1677
New York, NY, USA 10029

Tel:+1 212-659-8639
--