Sweet potato feathery mottle potyvirus (C1 isolate) virion and RNA purification.
A procedure for the purification of a Peruvian isolate (C1) of sweet potato feathery mottle potyvirus (SPFMV) and infective RNA has been developed. The use of Hepes [N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid] buffer containing urea and sodium EDTA as a base for tissue extraction and virus suspension enabled good yields of virus (35-50 mg/100 g) to be obtained from Nicotiana benthamiana L. Domin. A short RNA isolation procedure yielded infectious RNA, from which ds cDNA of nearly genome size could be obtained. Sweet potato feathery mottle potyvirus, Purification, RNA isolation, cDNA synthesis.
Nakashima
J T
JT
International Potato Center (CIP), Pathology Department, Lima, Peru.
Salazar
L F
LF
Wood
K R
KR
D002021:Buffers; D018076:DNA, Complementary / Q000096:biosynthesis; D004492:Edetic Acid; D006531:HEPES; D010936:Plant Extracts; D010947:Plants, Toxic; D017800:Potyvirus / Q000254:growth & development / Q000302:isolation & purification* / Q000472:pathogenicity / Q000648:ultrastructure; D012367:RNA, Viral / Q000302:isolation & purification*; D014026:Nicotiana / Q000382:microbiology*; D014508:Urea; D014675:Vegetables / Q000382:microbiology; D014771:Virion / Q000302:isolation & purification* / Q000648:ultrastructure; D014776:Virus Cultivation
D002021:Buffers; D018076:DNA, Complementary; D010936:Plant Extracts; D012367:RNA, Viral; D014508:Urea; D004492:Edetic Acid; D006531:HEPES