transformation of meningococci

Joel Gagliardi jg121 at UMAIL.UMD.EDU
Thu Apr 23 09:42:08 EST 1998


In partial answer to your question - you may have some difficulty.  I
worked with Dan Stein and I can tell you that N. gon. has no fewer than 8
restriction and modification systems. Dan was working on a strain with
inactivated restriction enzymes but active methylases for use in preparing
plasmids for insertion to other strains.  You can reach him at
dstein at microb.umd.edu.

----------
> From: Alexey Merz <alexey at webcom.com>
> To: methods at net.bio.net
> Subject: Re: transformation of meningococci
> Date: Wednesday, April 22, 1998 4:48 PM
> 
> Dr. Kris Partridge wrote:
> > 
> > I'm about to try to produce meningococci that express a foreign
> > protein.  Does anyone know if meningococci will accept a plasmid?  To
> > the best of my knowledge, I've seen nothing in the liturature.
> > 
> > Thanks for your help,
> > 
> > Kris Partridge
> 
> Contact Dr.Xavier Nassif at: nassif at necker.fr
> 
> He's built a series of shuttle vectors (the pMGC series) with a variety
> of features. They can be mated into gonococci or meningococci. These
> are derived from pLES2 (Stein & al., 1983, Gene 25:241). The most
> useful pMGC vectors for your purposes are probably MGC10 (ErmR, with
> blue/white selection in E. coli) and 18.1 (KmR, smaller size, no 
> blue/white). 
> 
> For mating proceedures try:Seifert & So, _Methods in Enzymology_,
204:342.
> 
> We have these vectors too, but Xavier actually made them. 
> 
> I'll warn you in advance: they are finicky, and it can be hard to 
> work with them even in E. coli.
> 
> Alternatively, it's really easy to drop things into the Neisserial 
> chromosome - ideas on how to do this are suggested in the Seifert/So
> article. Best of luck...
>
.........................................................................
> Alexey Merz | URL: http://www.webcom.com/alexey | email:
alexey at webcom.com
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