New Vectors for Chromosomal Integration Enable High-Level Constitutive or Inducible Magnetosome Expression of Fusion Proteins in Magnetospirillum gryphiswaldense

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  Optimization strategy of (inducible) expression cassette (Fig. S1); sequence alignment of nonoptimized egfp and Magnetospirillum-optimized egfp (magegfp) using CLUSTALW (Fig. S2); fluorescence of Magnetospirillum-optimized GFP (MagEGFP) versus nonoptimized GFP (EGFP) expressed from the P_mamDC45 promoter (A), and Western blot of whole M. gryphiswaldense cells expressing EGFP or MagEGFP under the control of P_mamDC45 (B) (Fig. S3); fluorescence micrographs of M. gryphiswaldense ΔC strains carrying a chromosomal insertion of P_mamDC45-mamC-magegfp (JH1), P_tet-mamC-magegfp (JH2) induced with 70 ng ml^-1 Atet or uninduced, or P_mamDC45-mamC-egfp-magegfp (JH3) expression cassette (Fig. S4); fluorescence and DIC micrographs of isolated magnetosomes from M. gryphiswaldense MSR-1 ΔC, JH2 (induced), JH1, and JH3 (Fig. S5); transmission electron micrographs of M. gryphiswaldense SB6 expressing MagEGFP chromosomally from P_mamDC45, and optical density (OD₅₆₅) after overnight growth of strains SB6, SB7, and SB8 in comparison to the wild type (Fig. S6); quantitative Western blot of GFP standard curve and MM samples from strains JH1, JH2, and JH3, and corresponding GFP protein concentrations of those strains (Fig. S7); plasmids used in this study (Table S1); strains used in this study (Table S2); primers used in this study (Table S3); insertion sites of expression cassettes in M. gryphiswaldense strains (Table S4).

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