Figure 2. MMC Gene Expression and Structure.

(A) Fluorescent detection of nuclear-localized DsRed in MMC1 maize leaf; size bar, 50 μm.

(B, C) Detection of DsRed sectors in a T2 plant leaf from event V-1 under (B) bright-field and (C) fluorescence microscopy; size bars, 0.5 mm.

(D) high magnification view of image shown in (C) with the corresponding sector, comprising all cell layers, indicated by an asterisk; the edge of a sector that comprises only the adaxial cell layer is indicated by arrowheads, cells with typical DsRed expression are indicated by arrows. Size bar, 50 μm.

(E) MMC consisting of a pCHR758 backbone and a centromere-derived insert, gene expression cassettes (grey), centromeric inserts (box), BglII restriction sites (arrowheads), and probes used for FISH and Southern blot analyses are indicated.

(F) Southern blot of DNA digested with BglII and hybridized to probes 1–6 (E); Bands 1–4 measure 2,067, 3,167, 5,227 and 790 bp, respectively; those hybridizing to probes 5 and 6 vary in size, depending on the location of BglII sites within the centromeric DNA insert. MMC1 Control (c, lanes 1 and 5) DNA was purified from E. coli and hybridization patterns were compared to DNA from plant cell extracts derived from MMC1 events V-1 (lanes 2–4), Q-2 (lane 6), and V-4 (lane 7), as well as from plants transformed with pCHR758 (lane 8) and untransformed wild type (H99, lane 9). For events V-1 and Q-2, bands differing from bacterial grown controls are indicated (arrows and asterisk, respectively).