Transmating: conjugative transfer of a new broad host range expression vector to various Bacillus species using a single protocol

Table 2 sfGFP fluorescence of different bacterial species carrying pBACOV-sfGFP relative to B. subtilis RIK1285 pBACOV-sfGFP

Strains were grown for 24 h at 37 °C, 180 rpm (B. mycoides: 72 h at 30 °C, 180 rpm) in SM-Cas medium with supplements, if appropriate. For details, see Material & Methods. Fluorescence was detected with cultures diluted to OD₆₀₀ = 1 and the fluorescence intensity (excitation: 470 nm, emission 520 nm) and the OD₆₀₀ were determined. The relative fluorescence (fluorescence intensity/OD₆₀₀) was calculated by normalizing the measured fluorescence intensity by the OD₆₀₀. The relative fluorescence of B. subtilis RIK1285 pBACOV-sfGFP was defined as 100%. The given values are mean values ± standard deviation of triplicate measurements. The asterisk * indicates that the B. pumilus wild-type culture did not reach an OD₆₀₀ of 1 within the 24 h of incubation and therefore was not diluted for fluorescence measurements. n. d.: not detectable (defined as relative fluorescence < 0.7%)

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