Recombinant Virus!!

[ANNALISE at medmicro.uct.ac.za]

> From:          wen at zeus.ahabs.wisc.edu

>
> I am interested in producing a recombinant DNA virus by transfecting
> plasmid (carrying a selectable gene flanked by viral sequences) into
> virally infected cells.  Could anybody help me begin to find out about
> other systems (preferably for eukaryotic viruses) that use a similar
> method to introduce such genetic  changes?

Poxvirus recombinants are also made using this method.  There are a
number of genes that have been used for positive selection - gpt,
neo, hygromycin.  Also beta-galactosidase has been used a
reporter gene to identify recombinants. Besides homologous
recombination to make recombinants in vitro ligation is also a
suitable method to make recombinant poxviruses -  as long as there is
a suitable helper virus to rescue the recombinants (naked poxvirus
DNA is not infectious).
>
Good luck

Anna-Lise Williamson


>Dr Anna-Lise Williamson
Dept Of Medical Microbiology
University of Cape Town Medical School
Observatory 7925
Cape Town
South Africa
Fax:27-21-4066205