Figure 2
Acidothermus cellulolyticus endocellulase AcCel5A expression construct, AcCel5A Western blot, and enzymatic saccharification. (a) The backbone vector pCAMBIA1305.2 (www.cambia.org) has a glycine-rich protein (GRP) signal peptide for extracellular targeting. The catalytically active AcCel5A gene (see supplemental material 1) or AcCel5A catalytic dead gene (AcCel5A-D) replaced the catalase intron-GusPlus gene cassette and the terminator was nopaline synthase (nos) polyA. See Materials and Methods for details. (b) Western blot of wild-type empty vector control (EV), AcCel5A and AcCel5A catalytically inactive (AcCel5A-D) transgenic plant lines showing the presence or absence of the AcCel5A protein in plant stem tissue, and a standard of purified AcCel5A. (c) Enzymatic saccharification. AcCel5A expression in plant cell walls increases the % glucan conversion by approximately 25% over control levels. The AcCel5A-D data is from line 37-4-a and the AcCel5A data is from line 21-2-b.
