Molecular cloning, sequencing, and expression in Escherichia coli of the potato virus y cytoplasmic inclusion gene

Ohshima, K.; Inoue, A.K.; Shikata, E.

Archives of Virology 128(1-2): 15-27

1993


ISSN/ISBN: 0304-8608
PMID: 8418790
DOI: 10.1007/bf01309785
Accession: 002656944

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Abstract
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.

Molecular cloning, sequencing, and expression in Escherichia coli of the potato virus y cytoplasmic inclusion gene