Figure 2.
Identification of the translational start site of mCherry in M. tuberculosis. Plasmids carrying mCherry were electroporated into M. tuberculosis and transformants selected on solid medium using hygromycin. The predicted proteins expressed from each plasmid are - pCherry29 = mCherry226; pCherry30 = mCherry219; pCherry0 (control plasmid) = no mCherry expression. (A) Transformant colonies. (B) Fluorescence was measured in liquid culture. Cultures were measured at Ex587/Em610 and results are expressed as relative fluorescence units (fluorescence/OD). Data are the mean and standard deviation from three independent transformants (C) Western analysis of protein expression in E. coli. Cell-free extracts were generated from transformants carrying plasmids and probed with anti-mCherry antibodies. Lane 1: no plasmid. Lane 2- pCherry10 (mCherry235). Lane 3- pCherry29 (mCherry226). Lane 4- pCherry30 (mCherry219). A non-specific band reacting with the commercial antibody was seen in all lanes, including E. coli lacking a plasmid.