Fig. 2
From: Codon-optimized DsRed fluorescent protein for use in Mycobacterium tuberculosis
Expression of functional DsRed. a Sequence alignment of mCherry and DsRed. Protein sequences were aligned using Clustal 2.1 [19]. b Activity of full length DsRed expressed from mycobacterial promoters. pBlazeC1-Phsp60; pBlazeC8-PrpsA; pBlazeC10-PG13. Recombinant M. tuberculosis was resuspended in 10 mM Tris pH 8.0 to an OD580 of 0.25, 0.10, 0.05 and 0.01 in 12 × 100 mm glass culture tubes. Fluorescence was measured at Ex/Em 558/583 nm. Data are the average ± SD of three cultures. c Plasmids were transformed into E. coli and cell-free extracts analyzed by Western blotting; 10 µg protein were subjected to SDS-PAGE, blotted onto PVDF membrane and visualized with anti-DsRed antibody. Lane 1—E. coli (no plasmid); Lane 2—pRed1; Lane 3—pRedA1; Lane 4—pRedB1; Lane 5—pRedC1; Lanes 6 and 7—pBlazeC1; Lane 8—pBlazeC8; Lane 9—pBlazeC10. The arrow indicates the size of the DsRed protein