mkennedy at chmeds.ac.nz
Tue Dec 7 21:36:47 EST 1993
In article <1993Dec6.213759.24631 at alw.nih.gov>, bernard at elsie.nci.nih.gov (Bernard Murray) writes:
> In article <CHMFnw.1uz at usenet.ucs.indiana.edu>, jgraham at bronze.ucs.indiana.edu (the End) writes:
>> In <1993Dec5.205157.162 at sci.wfeb.edu> vaughan at sci.wfeb.edu writes:
>> >Has anyone had a positive experience with the Lac-Switch system
>> >sold by Stratagene? I am scaling up to try it, but have been
>> >burned before.Any comments welcome (relative to Lac Switch that is)
> Stratagene Catalogue 1993 - New Product -page 43
> LacSwitch Inducible Mammalian Expression System
> Basically it seems to use LacI to repress transcription and this can be
> released by treatment with IPTG (this all happening in mammalian cells).
> This system had me intrigued as well and I was hoping someone would take
> the plunge and let us know how it performs. Does anyone know of any
> equivalent system (commercial or non-commercial)?
> Bernard Murray, Ph.D.
Sounds very much like something described by Baim et al (1991) PNAS 88,
5072-5076: A chimeric mammalian transactivator based on the lac repressor that
is regulated by temperature and IPTG.
Uses a minimal SV40 promoter linked to lac operator sequences, and a lacI
activator protein containing the transcriptional activator domain of HSV VP16.
The chimeric lacI protein (LAP267) is heat sensitive at 39.5 degrees, but its
activity can be rescued by addition of IPTG. They report 1200 fold increases
in CAT activity 24hours after adding IPTG in this system.
I've never used it, and I'm not sure I really understand it - I just happended
to have a copy of the paper ;-}
NNNN NN Martin A Kennedy (E-mail = mkennedy at chmeds.ac.nz) ZZZZZZZ
NN NN NN Cytogenetic and Molecular Oncology Unit ZZZ
NN NN NN Christchurch School of Medicine ZZZ
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