Construction of an infectious cDNA clone and gene expression vector of Tobacco vein banding mosaic virus (genus Potyvirus)

Tobacco vein banding mosaic virus (TVBMV, genus Potyvirus) mainly infects solanaceous plants and is of increasing economic importance in China. Here, we report sequence determination of the full-length 5′-untranslated region of TVBMV isolate HN39 and construction of an infectious clone. The resultant clone, pTVBMV, which was stabilized by introducing three introns in the P3 and CI-encoding regions, induced similar disease symptoms and accumulated similar titers of virus in plants of Nicotiana benthamiana, Nicotiana tabacum and N. rustica as the wild type HN39 isolate. Mutation of arginine to isoleucine (R182I) or aspartic acid to lysine (D198K) in HC-Pro alleviated the symptoms of pTVBMV significantly, indicating a role of the two amino acids in regulating virulence of TVBMV. The Aequoria victoriae gene for green fluorescent protein was inserted between the NIb and CP encoding regions of pTVBMV and expressed stably in the systemically infected N. benthamiana leaves, indicating suitability of pTVBMV for expression of foreign proteins in plants.

► The TVBMV genome sequence was completed with the first 25 nucleotides of 5′-UTR. ► A full-length infectious clone of TVBMV was constructed. ► Three introns were inserted to stabilize the infectious clone. ► The FRNK and CDN motifs of HC-Pro were found to affect virulence of TVBMV. ► The infectious clone of TVBMV was engineered for use as a gene expression vector.