The development of a subtype B AMPV reverse genetics system

A reverse genetics system is the only available method to introduce specific mutations into the AMPV genome. To date no system for the AMPV subtype B had been reported. In the system described, plasmids for N, P, M2, the polymerase and the full genome were constructed under the control of the T7 promoter. Producing intact clones containing the virus polymerase gene proved highly technically demanding due to the issue of them containing sequences toxic to bacteria, thus leading to sections of the gene being spontaneously deleted. Nonetheless, once all clones had been generated and transfected into Vero cells previously infected with a fowlpox recombinant virus expressing T polymerase, AMPV was recovered.