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Figure 4 | BMC Molecular Biology

Figure 4

From: Activation of mRNA translation by phage protein and low temperature: the case of Lactococcus lactis abortive infection system AbiD1

Figure 4

Effect of temperature on expression of the abiD1 TIRI : luxAB fusion. (A) Translation of abiD1 TIR I:luxAB fusion in L. lactis IL1403 cells at 30°C and 18°C. The background level of luciferase activity at 30°C and 18°C was measured in the presence of the control plasmid pJIM1715 [50]. Results are means of 5 independent experiments. Luciferase activity is shown in arbitrary light units (lux/OD unit at an OD600 of 0.4). (B) Expression of FLAG-tagged LuxAB at 30°C and 18°C. L. lactis cells carrying FLAG tagged abiD1 TIR I:luxAB fusion plasmid were grown to an OD600 of 0.4 at either 30°C or 18°C in M17 medium. Twenty mg of total cell protein were separated on an 8% SDS-PAGE gel. Western blot was developed with anti-FLAG tag M2 monoclonal antibody. The control panel shows cell lysates without specific FLAG-tagged protein. Molecular masses of standard proteins (Prestained Protein Marker, Broad Range, New England BioLabs) are indicated on the right. (C) Transcription of abiD1 TIR:luxAB fusion. The amount of abiD1 TIR I: luxAB transcript in L. lactis cells grown at 30°C and 18°C was quantified by ImageQuant software after dot-blot Northern hybridization and by quantitative RT-PCR. Values are means of 3–7 measurements, expressed in arbitrary units.

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