Overexpression of the Synthetic Chimeric Native-T-phylloplanin-GFP Genes Optimized for Monocot and Dicot Plants Renders Enhanced Resistance to Blue Mold Disease in Tobacco (N. tabacum L.)
Figure 1
Schematic map of the plant expression vector constructs pKM24-ibm8 and pKM24-ibm10 containing the synthetic tobacco Phylloplanin gene (GenBank accession no. AY705384) fused in-frame with GFP. Two genes (native T-phylloplanin and GFP) were fused in-frame in constructs pKM24-ibm8 and pKM24-ibm10 with linkers of 3 and 16 amino acids, respectively. The modified full-length transcript promoter (M24) of Mirabilis mosaic virus [2] directs the coding sequences of the respective native phylloplanin-GFP gene fusions. The chimeric gene sequence native T-phylloplanin-GFP was codon-optimized for both the dicot tobacco (pKM24ibm8; GenBank accession KF951257) and the monocot bent grass (pKM24ibm10; GenBank accession KF951258). A translational enhancer sequence (5′amv), the 35-nt long 5′-untranslated region of AlMV RNA 4, was fused with the gene. The apoplast targeting sequence (aTP) of the Arabidopsis 2S2 protein gene was fused in-frame with the coding sequence of native T-phylloplanin fused with GFP in the constructs. LT, left T-DNA border; RT, right T-DNA border; KanR, neomycin phosphotransferase II marker gene, and hygromycin resistance (HgR) directed by the nopaline synthase promoter (NosP), the 3′-terminator sequences (terminators) of the ribulose bisphosphate carboxylase small subunit (3′RbcS) and nopaline synthase (3′Nos) genes are also shown. The EcoRI, XhoI, SstI, NcoI, and ClaI restriction sites used to assemble these expression vectors are shown.
