age-
From: CCP4 bulletin board on behalf of ar...@xtals.org
Sent: Tue 5/5/2009 6:24 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
To clarify: I am not implying that I've worked with many proteins that
express better in XL1-blue cells than they
if it so then will i would be able to get good expression
> into this host???
>
> atul
> -Original Message-
> From: CCP4 bulletin board on behalf of ar...@xtals.org
> Sent: Tue 5/5/2009 6:24 AM
> To: CCP4BB@JISCMAIL.AC.UK
> Subject: Re: [ccp4bb] problem in tr
this host???
atul
-Original Message-
From: CCP4 bulletin board on behalf of ar...@xtals.org
Sent: Tue 5/5/2009 6:24 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
To clarify: I am not implying that I've worked with many proteins that
ex
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 da
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
@JISCMAIL.AC.UK
Sent: Monday, May 04, 2009 4:52 PM
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
Using pQE30, any E. coli is an expression host. Because it uses
the T5
promoter, you don't need an E. coli strain carrying the T7 RNA
polymerase
(so, you don't n
To: CCP4BB@JISCMAIL.AC.UK
Sent: Monday, May 04, 2009 4:52 PM
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
Using pQE30, any E. coli is an expression host. Because it uses the T5
promoter, you don't need an E. coli strain carrying the T7 RNA polymerase
(so, you don't n
alfa,but not in any of expression host
>
>
>
> From: ar...@xtals.org [mailto:ar...@xtals.org]
> Sent: Mon 5/4/2009 6:32 PM
> To: atul kumar
> Cc: ccp4bb@jiscmail.ac.uk
> Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
>
>
]
*Sent:* Mon 5/4/2009 6:32 PM
*To:* atul kumar
*Cc:* ccp4bb@jiscmail.ac.uk
*Subject:* Re: [ccp4bb] problem in transformation of pqe 30 clone
Hi,
You are using a pQE vector which has a T5 promoter. T5 is a native-like
promoter, recognized by the E. coli RNA polymerase - and this means that
even
able to
transform it into dh5 alfa,but not in any of expression host
From: ar...@xtals.org [mailto:ar...@xtals.org]
Sent: Mon 5/4/2009 6:32 PM
To: atul kumar
Cc: ccp4bb@jiscmail.ac.uk
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
Hi,
You are using a pQE vector which has a
m: ar...@xtals.org [mailto:ar...@xtals.org]
Sent: Mon 5/4/2009 6:32 PM
To: atul kumar
Cc: ccp4bb@jiscmail.ac.uk
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
Hi,
You are using a pQE vector which has a T5 promoter. T5 is a native-like
promoter, recognized by the E. coli RNA polymerase
To: atul kumar
Cc: ccp4bb@jiscmail.ac.uk
Subject: Re: [ccp4bb] problem in transformation of pqe 30 clone
Hi,
You are using a pQE vector which has a T5 promoter. T5 is a native-like
promoter, recognized by the E. coli RNA polymerase - and this means that
even with lac operatorsupstream there is a
Hi,
You are using a pQE vector which has a T5 promoter. T5 is a native-like
promoter, recognized by the E. coli RNA polymerase - and this means that
even with lac operatorsupstream there is a huge amount of leakage in this
system. If your protein is even moderately toxic then you have issues.
You
i have cloned my gene successfully into qiagen vector into pqe30 but i do
transformation of this into BL21,pLys,Rosseta,C41, i dont get any colonies,comp
cells are good other clones give good no of colonies upon transformation.
can someone help???
thanks
atul
-Original Message-
From: CC
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