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1 May 2012 Prokaryotic expression of a Thunberg fritillary mosaic virus CP gene and antiserum preparation
Wei Chuanbao, Wu Qiyao, Yang Yu, Liu Chunyun
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Abstract

Wei, C. B., Wu, Q. Y., Yang, Y. and Liu, C. Y. 2012. Prokaryotic expression of a Thunberg fritillary mosaic virus CP gene and antiserum preparation. Can. J. Plant Sci. 92: 495–500. Fritillariaanhuiensis is a valued traditional Chinese medicinal plant. Thunberg fritillary mosaic virus (TFMV) was found to cause mosaic symptoms. Virus particles of the Lu'an isolate were purified and its RNA genome was extracted. cDNA was synthesized from the RNA and specific PCR primers were used to amplify the virus coat protein (CP) gene. It shared 96.8% nucleotide identity and 97.6% amino acid identity with the sequences of CPs from the other TFMV isolates. The CP gene was inserted into pSBET and expressed in Escherichia coli BL21 (DE3) plys S strain. The expressed protein was first purified by 12% (wt/vol) SDS-PAGE and subsequently by 5–20% (wt/vol) gradient SDS-PAGE. Antiserum against the CP was raised in mouse and its specificity was confirmed by Western blot analysis suggesting that it may be useful for detecting the virus.

Wei Chuanbao, Wu Qiyao, Yang Yu, and Liu Chunyun "Prokaryotic expression of a Thunberg fritillary mosaic virus CP gene and antiserum preparation," Canadian Journal of Plant Science 92(3), 495-500, (1 May 2012). https://doi.org/10.1139/CJPS2011-138
Received: 12 January 2011; Accepted: 1 December 2011; Published: 1 May 2012
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KEYWORDS
antiserum preparation
coat protein gene
dépistage des virus
expression par un procaryote
Fritillaria anhuiensis
Fritillaria anhuiensis
gène CP
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