(NONE)

Warren Gallin wgallin at gpu.srv.ualberta.ca
Wed May 3 22:41:02 EST 2000



"Hiranya S. Roychowdhury" wrote:

> I have no knowledge of such a size clonable in a phagemid/plasmis. But, if
> such a thing exists for the usual host cells, I'd like to know too, please.
>
> At 05:32 PM 5/3/00 +0100, yan wu wrote:
> >I am looking for plasmid or phagemid vector(s) which
> >can hold 50kb DNA insert. I
> >appreciate it very much if anybody can send me
> >information about the name of the plasmid
> >and the place I can get it. Thank you very much. My

For that size people have typically used cosmids.  The only difference between
cosmids and plasmids is the inclusion of the lambda cos site in the vector,
allowing packaging into a lambda phage particle for easy transfection of the
bacteria.  The high transformation rate is mainly important for making
libraries.
    However, why not clone into your plasmid of choice and electroporate to
transfect your bacteria.  It may not be the most efficient method, but if you
are not making libraries it probably won't matter.
    As far as I know there is nothing special about plasmids that prevents
them from holding a big insert.  The reason people haven't used them for that,
historically, is that the chemical transfections are size-limiting.  I don't
think electroporation is.

Warren Gallin
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