Journal of General Plant Pathology (2008) 74, 97-100

Hideki Yamamoto and Shin-ichi Fuji (2008)
Rapid determination of the nucleotide sequences of potyviral coat protein genes using semi-nested RT-PCR with universal primers
Journal of General Plant Pathology 74 (2), 97-100
Abstract: A rapid method (<20 h) for determining the nucleotide sequences of potyviral coat protein (CP) genes was developed. The procedure involves reverse transcription using total RNA from a potyvirus-infected plant, followed by polymerase chain reaction (PCR) with the universal primers Sprimer/M4. The expected fragment, purified from an agarose gel, is used in semi-nested PCR with three degenerate primer sets: Sprimer/D1000, Sprimer/RYAFDFY-C and U341/M4. The three purified fragments are directly sequenced using the respective primers D1000, RYAFDFY-C and U341, then the complete CP sequence is assembled. All 11 potyviruses tested were efficiently sequenced by this method.
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Link to article at publishers website
Database assignments for author(s): Shin-ichi Fuji

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

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Pest and/or beneficial records:

Beneficial	Pest/Disease/Weed	Crop/Product	Country	Quarant.
	Potyvirus (genus)