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
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.
MCB graduate program
University of Wisconsin