RCNMV replicase
RCNMV replicase products are (+)-
stranded
Photographs and illustrations can be viewed by clicking the highlited texts
- Northern hybridization of in vitro transcripts of RCNMV using RCNMV replicase products as
probe. The 32P-labeled replicase products were produced by F3 fraction replicase in the absence of
exogenous templates. After phenol-chloroform extraction and ethanol precipitation, the replicase
products were used to hybridize with RCNMV transcripts of both (+)- and (-)-strand. The (+)- (lane 2)
and (-)-strand (lane 6) of RCNMV RNA-1 were synthesized by T7 and T3 RNA polymerases from
pFRC11 (Xiong and lommel, 1989), respectively. The (+)- (lane 3) and (-)-strand (lane 5) of RCNMV
RNA-2 were synthesized by T7 and T3 RNA polymerases from pBS1085 (Xiong and Lommel, 1991),
respectively. Lane 1 is BRL 1 kb DNA standards. Lane 4 contains linearized pBluescript plasmid
(stratagene) as a control. Panel A: Ethidium bromide stained RCNMV transcripts after fractionation on
a 1.2% agarose gel. Panel B: Autoradiograph of the Northern blot. The film was intentionally over-
exposed (6 days) to show the extremely weak hybridization of the replicase products with the (+)-
strand RCNMV transcripts. Almost all the replicase products hybridized with the (-)-strand transcripts,
indicating that the replicase products are (+)-stranded.
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