In article <wen-0401951919300001 at acplab.ahabs.wisc.edu> wen at zeus.ahabs.wisc.edu writes:
>>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?
We work with Adenovirus and have several systems for inserting genes
into the viral genome. One is the commonly used system developed by
Frank L. Graham (a medline search for his name should turn up at least
100 useful articles). Another is to CaPO4-transfect human 293 cells
with adenoviral DNA and a plasmid containing at least 1KB of homologous
sequence flanking the selectable gene of interest. This method needs
to direct the insertion into the E3 region of the virus, as this region
is not required for viral replication in vitro. With various deletions
of the E3 region, you can insert as much as 4KB of exogenous sequence.
Albert Einstein College of Medicine, NY