Figure 1.

Schematic representation of multicistronic expression constructs for epidermis-specific engineering of sesquiterpene formation. Two expression constructs were designed within the T-DNA, indicated by the left (LB) and right (RB) borders, of the binary pMCS vector. Both synthetic expression constructs are put under the control of an AtCER5 promoter sequence and inserted between the gateway attachments sites (attB1 and attB2). Each of the multicistronic expression constructs contains the coding sequences for three proteins: a prenyl transferase (AtFPPS or ShzFPPS), a terpene synthase (ShTPS12 or ShSBS), and enhanced green fluorescent protein (eGFP). The three individual coding sequences within both multicistronic expression constructs are linked by a short nucleotide sequence encoding the self-processing foot-and-mouth disease virus 2A oligopeptide (F2A). Other elements located within the T-DNA are the octopine synthase terminator (OCS-t), mannopine synthase promoter (MAS-p), and terminator (MAS-t) and phosphinothricin acetyltransferase (BlpR). The bars above the pC5-FTG and pC5-zFSG constructs indicate their size (in base pairs) and the location of the three primer pairs used for RT-PCR analysis of AtFPPS, ShzFPPS, and eGFP expression.