Construction of pJL22/FoMV, the full length FoMV construct. (a) Source in vitro transcription construct of native FoMV . (b) Recipient binary vector, pJL22 . A PCR fragment with a phosphorylated blunt upstream end and a NotI site at the downstream terminus was created by PCR from pFoMV10. This was ligated (1) into the StuI (blunt) and NotI sites of pJL22 to create pJL22/FoMV5'. (c) Next, the remainder of the FoMV sequence was added by creating a restriction fragment from pFoMV10 with PmlI/XbaI digestion. This was ligated (2) to the same sites in pJL22/FoMV5' to create pJL22/FoMV (Figure 3).