Dr. J.P. Clewley (jclewley at crc.ac.uk) wrote:
> One thing I forgot to mention is that its more difficult to engineer
> a negative strand virus than a positive strand one. If its +ve, you
> can make a cDNA clone & transfect it in cells & recover progeny virus.
> Works for alphaviruses (eg. Sindbis) picornaviruses (e.g. polio),
> retroviruses (e.g. HIV). Recovering -ve strand virus from transfected
> DNA has not been done reproducibly, as far as I know. Is there a
> confirmed example I'm missing?
We generate infectious clones of the NADL-2 strain cloned in pUC19 by
transfection. The virus is produced even if the vector is double stranded. We
are currently doing this with other strains as well. Furthermore, this enables
me to generate chimeric viruses and mutants in pUC and then producing the
virus by transfection (paper to be submitted shortly... film at eleven!).
Benoit_Hebert at iaf.uquebec.ca
Virology Research Center
Laval, Quebec, Canada H7N 4Z3
phone: (514) 687-5010, ext. 4636
fax: (514) 686-5626