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Cloning, sequencing, and expression inEscherichia coli of the coat protein gene of a new potyvirus infecting South AfricanPassiflora

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Summary

Complementary DNA representing 1418 nucleotides (nt) of the 3′-poly(A)-proximal tract of the genomic RNA of a potyvirus causing woodiness disease in South African passionfruit, was cloned and sequenced. The sequence contained a single long open reading frame (ORF) of 1188 nt with no initiation codon, and a 3′-non-coding region (3′-NCR) of 230 nt followed by a poly-adenylate tract. Comparison of the ORF with other potyviral coat protein (CP) sequences led to the prediction that a 279 residue CP of MW 31722 is encoded by 836 nt at the 3′-terminus of the ORF. This virus is not merely a South African strain of passionfruit woodiness virus (PWV): the deduced CP sequence is only distantly related to CPs of other sequenced strains of PWV, although it is part of a distinct subgroup of potyviruses related to PWV. The virus was therefore designated as South African passiflora virus (SAPV). The DNA containing the putative CP was cloned into the pUEX 2 bacterial expression vector and expressed inEscherichia coli as a β-gal-CP fusion protein. The fusion protein reacted with polyclonal antisera raised against the native virus, and antisera raised against partially purified fusion protein reacted with viral CP in Western blots.

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Author notes

  1. R. J. Brand

    Present address: Fruit and Food Technology Research Institute, Stellenbosch, South Africa

  2. J. T. Burger

    Present address: Plant Biology Research Centre, Pretoria, South Africa

Authors and Affiliations

  1. Department of Microbiology, University of Cape Town, Rondebosch, South Africa

    R. J. Brand, J. T. Burger & E. P. Rybicki

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  3. E. P. Rybicki
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Brand, R.J., Burger, J.T. & Rybicki, E.P. Cloning, sequencing, and expression inEscherichia coli of the coat protein gene of a new potyvirus infecting South AfricanPassiflora . Archives of Virology 128, 29–41 (1993). https://doi.org/10.1007/BF01309786

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  • DOI: https://doi.org/10.1007/BF01309786

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