Recombinant Virus!!

[wen at zeus.ahabs.wisc.edu]

Dear Virologists!

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?

I am aware that recombinant Baculoviruses are commonly produced by
transfection/infection.  Is there anybody reading this that has experience
with this method?  I am interested to know:

1.)  At what frequency do recombinants appear?

2.)  Do you think that cell type will drastically effect recombinant
frequency?  (ie: can I assume that data from one system is in the same
ballpark?).

3.)  Does any particular transfection method  (electroporation,
liposome-mediated, CaPhosphate, or others) work better than others?

4.)  How much homologous DNA seems to be necessary to acheive an
observable recombinant frequency?  

Comments and suggestions are gratefully welcomed.  E-mail is fine
(evans at ahabs.wisc.edu), but I also read this bulletin board religiously.

Thanks,

Pete Evans
MCB graduate program
University of Wisconsin