Fig. 1

One-step assembly of a Potyvirus infectious clone by a home-made Gibson assembly enzymatic premix. a Genomic organization of tobacco vein mottling virus (TVMV) and cloning strategy. Three cDNA fragments spanning the TVMV genome were assembled into a linearized T-DNA binary vector (pLX backbone); the PCR primers used are indicated (red arrows). b Digestion profiles are shown of purified plasmids from selected Escherichia coli colonies. Left, DNA size marker; right, the XbaI-digestion pattern computed for pLX-TVMV is shown (**, a TVMV-specific fragment). c Diagram of pLX-TVMV, a binary vector that includes a cDNA copy of the TVMV genome and regulatory sequences for its plant delivery and expression. Vector components are indicated (bottom); nptI, kanamycin resistance gene