Figure 6. Model for subcloning of a gene of interest into a library of compatible expressions vectors.
(A) A gene of interest can be cloned as a single fragment in one entry construct (EC) or cloned as separate fragments in several entry constructs (ECs). BsaI sites 1 to 4: B1–B4. Example of three expression vectors consisting of a viral provector (V1), a TMV-based viral vector (V2) and a standard non-replicating expression vector (V3), and of the expression constructs obtained after cloning (E1–3). GOI, gene of interest; pol, TVCV RNA-dependent RNA polymerase; m, movement protein; SP, signal peptide; P, promoter. (B) Overview of the cloning strategy from entry construct to expression vector. Boxes flanking the LacZ fragment (blue) represent elements specific to the various expression vectors.