cosmid vectors

[Robert Kuhn]

> > 
> > We currently have several problems related to transforming Agrobacterium
> > for complementation.  First we have a rather large region which we would
> > like to transform which is not represented in at least two cosmid
> > libraries.  We have therefore been trying to subclone lambda clones (size
> > 16-20 kbp) from the region into transformation vectors. Has anyone else had
> > to go this route? Unfortunately we don't yet have the ideal vectors for
> > this.  Does anyone have a transformation vector with an SfiI site in it? 
> 
> Susan, 
> 
> We have a vector with a Cla site between borders (pLZO3) and a transformation 
> system whereby you could put your cosmid (~15-20 kb) directly into our Agro 
> strain.  It requires packaging your cosmid product and infecting our 
> lambda-receptive strain.  It would also take a little engineering if you MUST 
> use the Sfi site.  See G.R. Lazo, et al.  Biotechnology 9:963-967.  Good luck.
> 
> 			--b0b kuhn 
> 
> > Or better yet does anyone have a cosmid vector with an SfiI site that will
> > accept and package 16kbp inserts?  We currently have pOCA18 and pCIT30,
> > both of which are not ideal for our purposes.
> >                     Thanks for help with any of these questions,  
> >                                    Susan Varnum        
> > 
> 
> ----------------------------------------------------------------
> Robert Kuhn, PhD              "Twenty years of schoolin'
> Sinsheimer Laboratories        and they put you on the day shift
> University of California    
> Santa Cruz, CA 95064  USA      (brown shoes don't make it)"
>                                            -F. Zappa (RIP) 
> email: rkuhn at orchid.ucsc.edu
> ================================================================
> 
>