Re: [ccp4bb] problem in transformation of pqe 30 clone

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

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Subject: Re: problem in transformation of pqe 30 clone
From: Ho-Leung Ng holeung {- at -} BERKELEY {- dot -} EDU
Date: 2009-05-04

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Subject: Scientists position - Australian Synchrotron
From: Sonya Sivaraj Sonya {- dot -} Sivaraj {- at -} SYNCHROTRON {- dot -} ORG {- dot -} AU
Date: 2009-05-04

Subject: Re: problem in transformation of pqe 30 clone
From: artem {- at -} XTALS {- dot -} ORG artem {- at -} XTALS {- dot -} ORG
Date: 2009-05-04

To clarify: I am not implying that I've worked with many proteins that
express better in XL1-blue cells than they would express in BL21(DE3) or
such if cloned into a pET vector or similar. In fact I can probably recall
only one or two that expressed *better* in XL1 cells. However in the good
old days pQE series of vectors was quite commonly used and I had things in
that were inherited from others - these 'things' were fairly simple to
express in XL1-blue whereas they gave me loads of trouble in other strains
and I was too busy/lazy to re-clone them.

Artem

> Hello Artem,
>
> We express almost all our proteins in BL21 derivatives. It sounds
> like you've worked with many proteins that express/behave better in
> XL1-Blue?
>
>
> ho
> UC Berkeley
>
> -------------------------------------------------------------------------------------------------
> XL1-Blue is a strain of E. coli. Whether it is or isn't an expression host
> depends on the definition, and I am not going to argue semantics.
>
> The T5 promoter works with regular garden variety RNApol of E. coli.
> Therefore ANY E. coli strain is an 'expression host' for vectors that
> contain this promoter.
>
> I've expressed many proteins in XL1-Blue and I see no reason why you can't
> express yours, either.
>
> Artem
>

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Re: problem in transformation of pqe 30 clone
From: Ho-Leung Ng holeung {- at -} BERKELEY {- dot -} EDU
Date: 2009-05-04

Next message:
Subject: Scientists position - Australian Synchrotron
From: Sonya Sivaraj Sonya {- dot -} Sivaraj {- at -} SYNCHROTRON {- dot -} ORG {- dot -} AU
Date: 2009-05-04