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Expression of Recombinant Potato leafroll virus Structural and Non-structural Proteins for Antibody Production

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Vector pMPM-A4Ω and vectors pQE-30 and pET-45b(+) containing the 6x His-tag sequence were used for expression of Potato leafroll virus (PLRV) structural and non-structural proteins in Escherichia coli. Coat protein (CP) and RNA-dependent RNA polymerase (RdRp)–fragments RdRp43-616 and RdRp304-537 were chosen for expression. A high level of CP and RdRp304-537 was obtained only in an expression system using pET-45b(+) vector and E.┬ácoli Rosetta-gami 2(DE3) cells. After purification, the His-tagged PLRV proteins were used for immunization of rabbits.

Keywords: Potato leafroll virus; antibody production; recombinant viral antigen

Document Type: Short Communication

DOI: http://dx.doi.org/10.1111/j.1439-0434.2010.01740.x

Affiliations: 1: Authors’ addresses: Institute of Experimental Botany, v. v. i., Academy of Sciences of the Czech Republic, Na Karlovce 1a, 160 00 Prague 6, Czech Republic 2: Potato Research Institute, Havlickuv Brod, Dobrovskeho 2366, 580 01 Havlickuv Brod, Czech Republic (correspondence to N. Cerovska. ), Email: cerovska@ueb.cas.cz

Publication date: February 1, 2011

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