Construction of an infectious full-length cDNA clone of potato virus M.
An infectious full-length cDNA clone of potato virus M (PVM) was produced. Total RNA was extracted from PVM-infected Nicotiana hesperis plants and used for cDNA synthesis. Subsequent RT-PCR produced two DNA fragments of about 5.5 and 3.2 kbp, which were ligated downstream of an enhanced 35S cauliflower mosaic virus promoter. After cloning of the enhanced 35S promoter with the PVM sequence into a modified pBIN19 plasmid and electroporation of Agrobacterium tumefaciens, the agroinoculated PVM full-length clone (pPVM-flc) led to systemic PVM infections in different host plants, causing symptoms indistinguishable from those caused by wild-type PVM.
Institute of Plant Diseases and Plant Protection, Leibniz Universitaet Hannover, Herrenhaeuser Strasse 2, 30419 Hannover, Germany.
S
S
Flatken
V
V
Ungewickell
W
W
Menzel
E
E
Maiss
D016960:Agrobacterium tumefaciens / Q000235:genetics; D017788:Carlavirus / Q000235:genetics*; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D018274:Electroporation; D023061:Gene Order; D010935:Plant Diseases / Q000821:virology; D010944:Plants / Q000821:virology; D012367:RNA, Viral / Q000235:genetics*
D018076:DNA, Complementary; D012367:RNA, Viral