Molecular cloning, sequencing, and expression in Escherichia coli of the potato virus Y cytoplasmic inclusion gene.
Complete nucleotide sequences of cytoplasmic inclusion (CI) genes of two strains of potato virus Y (PVY) were determined from six polymerase chain reaction (PCR)-amplified cDNA clones. The size of the CI genes of both ordinary (PVY-O) and necrotic strains (PVY-T13) was 1902 nucleotides, with a sequence homology of 83.4%. Comparison of the predicted amino acid sequences showed more than 90% homology. When these were compared with those of other potyviruses, the homology ranged from 53 to 61%. cDNAs of all or a part of the PVY-O CI gene containing an additional initiation codon (ATG) at the 5' end and a stop codon at the 3' end were constructed by PCR amplification and cloned into an Escherichia coli expression vector, pKK 223-3. Complete and truncated PVY-O CI proteins were successfully produced in E. coli as judged by reactivities with PVY-O CI protein-specific antiserum. To our knowledge, this is the first report on expression of PVY CI proteins in E. coli.
Ohshima
K
K
Department of Botany, Faculty of Agriculture, Hokkaido University, Sapporo, Japan.
Inoue
A K
AK
Shikata
E
E
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral; D004926:Escherichia coli; D005814:Genes, Viral; D007181:Inclusion Bodies, Viral; D008969:Molecular Sequence Data; D010942:Plant Viruses / Q000235:genetics*; D012328:RNA Viruses / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D011198:Solanum tuberosum; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D014764:Viral Proteins; C079046:cytoplasmic inclusion protein, Potato virus Y