Construction of a stable and highly infectious intron-containing cDNA clone of plum pox potyvirus and its use to infect plants by particle bombardment.
An infectious plum pox potyvirus cDNA clone was constructed placing a copy of the full-length sequence of the virus genome between an enhanced cauliflower mosaic virus 35S promoter and a nopaline synthase termination signal. Stabilization of the clone and faster growth of bacteria, in addition to higher plasmid yield, followed a modification consisting of the insertion of an intron which interrupted the viral open reading frame at the P3 region. This intron-containing clone was infectious when inoculated into plants after undergoing in vivo transcription and splicing. Particle bombardment delivery of the cDNA greatly increased the efficiency of plant infection.
López-Moya
J J
JJ
Centro Nacional de Biotecnología, Campus de la Universidad Autónoma, CSIC, Cantoblanco 28049, Madrid, Spain.
García
J A
JA
D003001:Cloning, Molecular; D018076:DNA, Complementary; D004279:DNA, Viral / Q000502:physiology*; D004926:Escherichia coli; D005818:Genetic Engineering; D007438:Introns; D016254:Mutagenesis, Insertional; D010947:Plants, Toxic; D017801:Plum Pox Virus / Q000235:genetics* / Q000472:pathogenicity; D012326:RNA Splicing; D014026:Nicotiana
D018076:DNA, Complementary; D004279:DNA, Viral