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Title
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Nucleotide sequence and in vitro expression of the capsid protein gene of tobacco ringspot virus.
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Authors
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B.Buckley,
S.Silva,
S.Singh.
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Ref.
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Virus Res, 1993,
30,
335-349.
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PubMed id
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Abstract
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The nucleotide sequence of the 3' terminal 2022 nucleotides (nt) of tobacco
ringspot virus (TobRV) RNA 2 has been determined. Protein microsequence analysis
of the amino-terminal residues of purified capsid protein localized the capsid
protein gene between nt 2014 and 583 (from the 3' terminus) of this sequence.
The proteolytic cleavage site that is processed to liberate the capsid protein
from the RNA 2-encoded polyprotein was identified as Cys-Ala. The predicted
translation product from the gene is a 477 amino acid long polypeptide with a
calculated MW of 53 kDa. The gene was modified at the 5' end to facilitate
sub-cloning, and to provide it with a methionine initiation codon. The modified
gene was sub-cloned, transcribed in vitro and expressed in a rabbit reticulocyte
lysate translation system, where it directed the synthesis of a 53 kDa
polypeptide. Garnier-Osguthorpe-Robson analyses of the secondary structure of
the capsid protein predicted the presence of three beta sheet domains, which
suggests that this nepovirus capsid may be structurally analogous to those of
the como- and picornaviruses. These and other results from computer analyses of
the nucleic acid and amino acid sequences, and comparisons with the capsid
proteins of nepoviruses and other related viruses are discussed.
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