Figure 7

Determination of enzyme amounts in crude extracts of XynA-, Xyn10A- and Xyn11B-expressing cv. I64 plants measured in mature green leaves (GL) and lower senescing leaves (SL) and a zymogram of the extracts. A. Three repeated extractions from GL and SL of the cv.I64 lines expressing XynA and Xyn10A from CEC4 and Xyn11B from CEC5 were analysed by western blot for quantification of recombinant proteins content. Each lane of XynA extracts from both GL and SL represents 2.5 mg of extracted leaf tissue; for Xyn10A and Xyn11B each lane represents 0.05 mg and 0.25 mg of extracted leaf tissue for GL and SL extracts, respectively. WT – extract of untransformed WT plants used as a negative control. Known amounts (in ng) of a c-myc-tagged control protein are indicated above the standard curve lanes. B. Zymogram of GL and SL extracts (combined of three repeats) for XynA, Xyn10A and Xyn11B, resolved on a 12% SDS-PAGE gel containing 0.1% (w/v) xylan. Each lane contains equivalent of 2.5 mg of extracted leaf tissue. Equal amount of extracts from WT leaves were used as a negative control. Arrows indicate expected sizes of the protein bands detected on western blots.