Archives of Virology (1997) 142, 1101-1116

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K.-S. Ling, H.-Y. Zhu, H. Alvizo, J.S. Hu, R.F. Drong, J.L. Slightom and D. Gonsalves (1997)
The coat protein gene of grapevine leafroll associated closterovirus-3: cloning, nucleotide sequencing and expression in transgenic plants
Archives of Virology 142 (6), 1101-1116
Abstract: A lambda ZAP II cDNA library was constructed by cloning cDNA prepared from a high molecular weight double-stranded RNA (dsRNA, ca. 18 kb) isolated from grapevine leafroll associated closterovirus-3 (GLRaV-3) infected tissues. This cDNA library was immuno-screened with GLRaV-3 coat protein specific polyclonal and monoclonal antibodies and three immuno-positive clones were identified. Analysis of nucleotide sequences from these clones revealed an open reading frame (ORF) which was truncated at the 3' end; the remainder of this ORF was obtained by sequencing a fourth clone that overlapped with one of the immunopositive clones. A total of 2028 bp was sequenced. The putative GLRaV-3 coat protein ORF, 939 bp, encodes a protein (referred to as p35) with a calculated Mr of 34 866. Multiple alignment of the p35 amino acid sequence with coat protein sequences from other clostero-viruses revealed that the consensus amino acid residues (R and D) of filamentous plant viruses are preserved in the expected locations. The GLRaV-3 coat protein gene was then engineered for sense and antisense expression in transgenic plants. Transgenic Nicotiana benthamiana plants that contain the sense GLRaV-3 coat protein gene produced a 35 kDa protein that reacted with GLRaV-3 antibody in Western blot.
(The abstract is excluded from the Creative Commons licence and has been copied with permission by the publisher.)
Link to article at publishers website
Database assignments for author(s): Kai-Shu Ling

Research topic(s) for pests/diseases/weeds:
molecular biology - genes


Pest and/or beneficial records:

Beneficial Pest/Disease/Weed Crop/Product Country Quarant.


Grapevine leafroll-associated virus 3