luciferase in vivo in mammals?
Michael Benedik
bchs1b at Elroy.UH.EDU
Fri Jun 10 15:07:51 EST 1994
In article <Pine.3.07.9406101546.A29092-a100000 at alpha.prospect.anprod.csiro.au>, davead at prospect.anprod.csiro.au (David Adelson) writes:
>
>Can anybody out there tell me if luciferase can be detected in vivo in
>mammalian cells or tissues? In cells this would be as a screen for
>expression and so would have to be non lethal as we want to pick the
>expressing cells and grow them up.
>
>Please post replies to me directly as my news reader is slow and unreliable.
>
>Thanks in advance,
> ------------------------------------------------------------------------
> | Dave Adelson E-mail: davead at prospect.anprod.csiro.au |
> | CSIRO Division of Animal Production * "All opinions are my own" |
> | Locked Bag 1, Delivery Centre * If you don't like my logic, |
> | Blacktown, 2148, AUSTRALIA * use your own. |
> |----------------------------------------------------------------------|
>
>
Yes.
firefly luciferase is what most people think of in the euk work because
it is a single gene product. However you need to break the cells open
to get in the substrate.
Bacterial luciferase has 2 subunits and therefore is tricky to express
in eucaryotes. However a number of groups have made gene fusios
fusions of the two subunits, and these work in both proks and euks.
Decanol will diffuse in without requireing cell lysis and hence this
can be used in vivo. But for use in mammalian cells see
Pazzagli et al 1992 Analytical Biochemistry 204:315-323.
----------------------------------------------------------------------
Michael Benedik INTERNET: Benedik at uh.edu
Dept. of Biochemical & Biophysical Sciences
University of Houston BITNET: Benedik at uhou
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