Appl Microbiol Biotechnol (2010) 87:1783–1789 1785 Fig. 1 Expression vectors constructed in this study. s.s. secretion signal sequence of tglA, N28 N-terminal 28 amino acid residues of Rhizopus oryzae lipase pro-region, mtglA mature region of tglA ACAACGCTCCCCTGAATGAGTTCCTCAGCGCT-3′ a n d 5 ′ - G G T T AT T T A A AT T T A C T T G T C AT C G TCATCCTTGTAGTCGTTCGCAGCCGCAACAGCA Fig. 3 GFP fluorescence micro- graphs of A. oryzae. A. oryzae/ GFP-CBM transformant (a, b, and c) A. oryzae/CBM-GFP transformant (d, e, and f). A. oryzae/GFP transformant (g, h, and i). Enlarged views in the circles in a, d, and g (b, e, and h, respectively). Proteinase K-treated transformants (c, f, and i). Each scale bar represents 50 μm Fig. 2 Western blot analysis of GFP, GFP-CBM, and CBM-GFP fusion proteins from cell wall fractions (lanes 1 to 3), cytosolic fractions (lanes 4 to 6), and supernatant fractions (lanes 7 to 9). Lanes 1, 4, and 7, A. oryzae/GFP-CBM transformant; 2, 5, and 8, A. oryzae/ CBM-GFP transformant; 3, 6, and 9, A. oryzae/GFP transformant A - 3 ′ o r 5 ′ - G G T T G C G G C C G C C T T G T C AT C G TCATCCTTGTAGTCGTTCGCAGCCGCAACAGCA A-3′. The amplified fragment was digested with SalI and SwaI, or SalI and NotI, respectively, and inserted into pISI-GFP-CBM. The resultant plasmids were designated as pISI-tglA and pISI-tglA-CBM, respectively. The DNA