Generation of infectious transcripts from Korean strain and mild mottle strain of potato virus X.
Full-length cDNAs of two different strains of Potato virus X (PVX-Kr and PVX-Mo) have been directly amplified by long template reverse transcription polymerase chain reaction (RT-PCR) using the 5'-end primer containing a SP6 or T7 RNA promoter sequence and the virus-specific 3'-end primer, and then constructed in plasmid vectors. Capped in vitro transcripts from cloned full-length cDNAs as well as those RT-PCR amplicons proved to be infectious systemically on tobacco plants. Symptom expression on tobacco plants from PVX-Mo transcripts was faster and severer than that from PVX-Kr. In replication stability test of transcripts derived from PVX clones, progeny viruses showed stable replication according to sequencing through passages. This highly infectious transcript system from the full-length cDNA clones for PVX can be useful for recombinant molecules for functional analysis of viral proteins in plant-virus interaction study as well as for expression of foreign protein in planta.
Choi
Sun Hee
SH
Plant Virus GenBank, Division of Environmental and Life Sciences, Seoul Women's University, Seoul, 139-774, Republic of Korea.
Ryu
Ki Hyun
KH
D036022:Capsid Proteins / Q000235:genetics; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000382:microbiology*; D017863:Potexvirus / Q000235:genetics* / Q000472:pathogenicity*; D012367:RNA, Viral / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014026:Nicotiana / Q000382:microbiology*; D014158:Transcription, Genetic
D036022:Capsid Proteins; D012367:RNA, Viral