The expression and secretion of the recombinant proteins in P. fluorescens . A. The intracellular expression and secretion of GFP-TliA and GFP-LARD3 in P. fluorescens cultured in LB were detected by western blotting using anti-LARD antibodies. The P. fluorescens cells expressing the designated proteins with (+) or without (−) TliDEF were isolated from two different extracellular media by double centrifugation. B. The same expression was performed in 2× LB (double concentrated LB). Lane 1, pDSK-TliA; lane 2, pDX-TliA; lane 3, pDX-GFP-LARD3; lane 4, pDX-GFP-TliA. The arrows indicate the sizes of GFP-LARD3 (38.2 kDa), TliA (49.9 kDa), and GFP-TliA (77.4 kDa). Notice the intrinsic TliA indicated by the arrows.