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. 2022 May 30;34(8):3028–3046. doi: 10.1093/plcell/koac153

Figure 2.

Figure 2

Cloning of CUE8. A, Map-based cloning of the CUE8 gene, AT5G22640, encoding TIC100/EMB1211. Upper, abbreviated name of the polymorphisms used for mapping, position along chromosome 5 (in kb), and number of recombinants at those positions identified in the indicated mapping populations. This identifies an 82-kb region, containing 19 open-reading frames (middle). A combination of strategies (Supplemental Table S2) identifies AT5G22640 (TIC100) as the CUE8 locus, whose exon/intron structure is shown. A point mutation (lower) results in a single amino acid substitution (G366R) in the TIC100 protein sequence. B, Complementation of cue8 with 35S:TIC100, carrying a TIC100 cDNA under the control of a 35S promoter. Plants shown before and after the selection of transformants. C, Diagnostic PCR confirming the presence of the 35S:TIC100 transgene in complemented plants. Positive (+ve) control, plasmid DNA harboring the construct. Negative (−ve) control, DNA from plant prior to transformation.