Figure 5.

Identification of the (SP4) ₁₈ ‐EGFP and (SP) ₃₂ ‐EGFP transgene products accumulated in root tissues. (a) SDS‐PAGE separation of (SP4)₁₈‐EGFP and (SP)₃₂‐EGFP products purified from the root tissues. Five μg each of the protein samples was run on a 4%–20% Tris‐HCl gel and stained with Coomassie blue; (b) N‐terminal peptide sequencing of the purified transgene products. ‘X’ denotes unidentified amino acid; (c) Anti‐EGFP Western blotting assay of the (SP4)₁₈‐EGFP or (SP)₃₂‐EGFP expression in hairy roots (HR) and E. coli. Ten μL of clarified root extracts (12‐day‐old culture) or 5 μL of E. coli lysates was loaded. Crtl⁺: EGFP standard (50 ng). (d) Anti‐EGFP Western blotting detection of the (SP)₃₂‐EGFP products within root tissues before and after β‐elimination reaction. Ten μL of clarified root extract (before treatment) or root extract equivalent (after treatment) was loaded.