Figure 3. Schematic representation of the PCR strategy used to clone an apple pgip gene.

A. pIPGIP which contains the ipgip fragment used to design InvPCR primers AP-PGIP-INV_L (INV_L) and AP-PGIP-INV_R (INV_R).

B. BglII InvPCR clone, pAppBglII. Location of the primers used for InvPCR (INV_L and INV_R) is indicated. The 98bp and 166bp overlaps between the BglII InvPCR clone and the ipgip fragment are shown by solid blue boxes.

C. NsiI InvPCR clone. The positions of the primers AP-PGIP-INV_L-2 (INV_L-2) and AP-PGIP-INV_R-2 (INV_R-2), designed to sequences from pAppBglII, are indicated. The 190bp overlap between pAppBglII and pAppNsiI is indicated by a solid red box.

D. The composite apple pgip gene constructed using sequences obtained from pIPGIP, pAppBglII and pAppNsiI. B, BglII; E, EcoRV; H, HindIII; N, NsiI; X, XbaI.