Prokaryotic expression of a Thunberg fritillary mosaic virus CP gene and antiserum preparation

摘要

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. Fritillaria anhuiensis 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.