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Chimeric Virus as a Source of the Potato Leafroll Virus Antigen

Skurat, Eugene V., Butenko, Konstantin O., Kondakova, Olga A., Nikitin, Nikolai A., Karpova, Olga V., Drygin, Yuri F., Atabekov, Joseph G.
Molecular biotechnology 2017 v.59 no.11-12 pp. 469-481
Potato leafroll virus, RNA, Tobacco mosaic virus, antibodies, coat proteins, codons, complementary DNA, genes, genetic engineering, intergenic DNA, viral antigens, virion, viruses
Large quantities of potato leafroll virus (PLRV) antigen are difficult to obtain because this virus accumulates in plants at a low titer. To overcome this problem, we constructed a binary vector containing chimeric cDNA, in which the coat protein (CP) gene of the crucifer infecting tobacco mosaic virus (crTMV) was substituted for the coat protein gene of PLRV. The PLRV movement protein (MP) gene, which overlaps completely with the CP gene, was doubly mutated to eliminate priming of the PLRV MP translation from ATG codons with no changes to the amino acid sequence of the CP. The untranslated long intergenic region located upstream of the CP gene was removed from the construct. Transcribed powerful tobamovirus polymerase of the produced vector synthesized PLRV CP gene that was, in turn, translated into the protein. CP PLRV packed RNAs from the helical crTMV in spherical virions. Morphology, size and antigenic specificities of the wild-type and chimeric virus were similar. The yield of isolated chimera was about three orders higher than the yield of native PLRV. The genetic manipulations facilitated the generation of antibodies against the chimeric virus, which recognize the wild-type PLRV.