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co-existence of expression vector and promoterless reporter vector inthe same cells

co-existence of expression vector and promoterless reporter vector inthe same cells - Protocols and Methods Forum

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  #1  
Old 04-04-2007, 03:04 PM
waikhay
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Default co-existence of expression vector and promoterless reporter vector inthe same cells



Hi all,

I am currently working on cloning and expressing a bacterial regulatory
protein using an expression vector to test for its binding to a bacterial
promoter which is cloned into a promoterless reporter vector that can
ideally co-exist with the expression vector in the same E. coli cells.
However, the expression vectors (e.g. pQE seris by Qiagen and pET series by
Novagen) and the promoterless reporter vectors (e.g. pZsGreen1-1 and
pDsRed-Express-1 by Clontech) that I found share the origin of replication
which falls into the same plasmid incompatibility group. Does anyone out
there have an idea of which expression and promoterless reporter vectors I
can use that can also co-exist in the same cell? Any form of help would be
greatly appreciated!!

Cheers,
Waikhay
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  #2  
Old 04-05-2007, 02:31 AM
Aawara Chowdhury
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Default co-existence of expression vector and promoterless reporter vector in the same cells

In <[Only registered users see links. ].net >,
waikhay <[Only registered users see links. ]> wrote:


pET and pQE vectors have derivatives of the ColE1 origin. Your reporter
plasmid can have the pSC101 origin, as it lies in a different incompatibility
group.

AC
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Email: echo 36434455860060025978157675027927670979097959886449 930P | dc
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  #3  
Old 04-05-2007, 03:11 AM
Aawara Chowdhury
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Default co-existence of expression vector and promoterless reporter vector in the same cells

In <8cZQh.6248$[Only registered users see links. ]>,
Aawara Chowdhury <[Only registered users see links. ]> wrote:


Also, if you don't have access to a plasmid with a pSC101 origin, I would
be happy to send you one - just write me at the email address below.

NEB also provides plasmids with the p15A origin, which is also compatible
with ColE1-origin plasmids. pACYC177 and pACYC184 are shipped for free
if requested along with an order.

AC
--
Email: echo 36434455860060025978157675027927670979097959886449 930P | dc
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  #4  
Old 04-09-2007, 09:50 AM
Trond Erik Vee Aune
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Default co-existence of expression vector and promoterless reporter vectorin the same cells

waikhay wrote:


For high expression in E.coli and other gram negative bacteria I can
recommend our own in-house broad-host expression vectors. They are based
on the RK2-replicon (IncP) and should therefore be compatible with most
commercial vector systems.

For more info:
[Only registered users see links. ]
[Only registered users see links. ]

Best regards,
Trond Erik

--
Trond Erik Vee Aune
Department of Biotechnology, NTNU
[Only registered users see links. ]

- Must be sad being a dyslectic, agnostic insomniac, lying
awake during the night, wondering if there really is a dog
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