Regulation of the tet promoter on pBR322????

[Matthew Nilles]

In article <30NOV199410073178 at watson.bms.com>, patel_p at watson.bms.com
(PRAMATHESH PATEL) wrote:

> Hi:
>     I have constructed a home made vector for a specific use.  The vector
> made vector which is essentially like the pUC vectors except that I have
> replaced the lac promoter with the tet promoter from pBR322.
>     The questions I have are:
> 
> 1) Is the tet promoter of pBR regulated by a tet repressor??
> 
> 2) If I "express" the tet repressor in the same cell will I be able to get
> a "regulateble" induction of tet promoter with chloro-tetracycline
> 
> 3)  Does anybody know where I could get the tet repressor gene.
> 
> Any comments or suggestions will be appreciated.
> 
> Thanks in advance
> Prem
> 
> 
> 
> e-mail:   Patel_P at BMS.com
Patel;

In response to your questions;

1) Yes, the tet promoter of pBR can be regulated by a repressor, although
the repressor is not present on pBR.  The repressor is called TetR.

2) Yes, expression of the repressor in the same cell should give
'regulateable' induction of the tet promoter, However if your plasmid has
a pUC copy number it may be difficult to get enough TetR made to repress
all copies of the plasmid.

3) A good source for TetR contructs is Kevin Bertrand (my major advisor)
here at Washington State University.

For more specific information E-mail me at:
KBERT at jaguar.csc.wsu.edu