BL21 for 35-S-Met labeling?

Miguel Zamora mzamora at bcm.tmc.edu
Fri Feb 18 19:25:42 EST 2000


Yoram,

I used BLR(DE3)pLysS for radiolabeling recombinant proteins, but BL21 should
do the trick.  In the basic protocol you induce the expression of T7 pol
with IPTG for about one hour,  inhibit transcription of bacterial genes with
rifampicin for about another hour, and then add the radiolabel and incubate
for another hour.  I used M9 media supplemented with 18 aa and thiamin.  I
have two or three references describing the protocol although not for
radiolabeling.  Give me a day or two to find them (we just moved to a brand
new lab and things are a bit messy here).

Cheers,

Miguel




Yoram Gerchman wrote:

> Dear Netters
> Did anyone try to use the E.coli BL21 strain for specific labeling of a
> protein with the T7 RNA polymerase?
> Regards Yoram
> .........................................................................
> "Support bacteria - they're the only culture some people have"
>
>                                      (anonymous)
>
> Yoram Gerchman
> Division of Microbial & Molecular Ecology
> The Institute of Life Sciences
> Hebrew University of Jerusalem
> Givat Ram, Jerusalem 91904, Israel
>
> ---
-------------- next part --------------
A non-text attachment was scrubbed...
Name: mzamora.vcf
Type: text/x-vcard
Size: 298 bytes
Desc: Card for Miguel Zamora
Url : http://iubio.bio.indiana.edu/bionet/mm/methods/attachments/20000218/b45b521d/mzamora.bin


More information about the Methods mailing list