EXPRESSION OF NUCLEOCAPSID PROTEIN OF CAPSICUM CHLOROSIS VIRUS IN A PROKARYOTIC SYSTEM AND ITS APPLICATION IN VIRUS DETECTION

X. Rao, Y. Li, Z. Wu, Y. Liu, W. Wang
doi: 10.4454/JPP.V96I2.032
Abstract:
Capsicum chlorosis virus (CaCV), which belongs to the Watermelon silver mottle virus (WSMoV) serogroup of the genus Tospovirus, can seriously harm economically important crops. A CaCV strain from Phalaenopsis orchids (CaCV-pha) was identified by RT-PCR. The amino acid sequence of the CaCV-pha nucleocapsid protein (NP) shared 92.0% to 97.1% identity with comparable proteins of other CaCV isolates, with which it was phylogentically related. Since the identification of CaCV-infected crops requires the availability of sensitive diagnostic tools, a prokaryotic expression system was used to obtain a recombinant CaCV-pha NP for antiserum production. The CaCV-pha NP gene was subcloned into the pET-28a (+) vector and transformed into Escherichia coli Rosetta (DE3) cells for expression. After purification, the recombinant protein was injected into rabbits to generate a polyclonal antiserum which, in Western blot tests reacted strongly with the recombinant protein used as antigen. The availability of an anti-CaCV-NP serum will facilitates field surveys for, and further research on CaCV.
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