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How Do Haloarchaea Synthesize Aromatic Amino Acids?

Figure 3

Schematic representations of a single cross-over event between the chromosomal DNA of H. salinarum R1 and plasmid pMG501.

P1,P2 and P3 and their associated arrows indicate the relative positions and orientations of the primers used for PCR. After in-frame integration of plasmid pMG501, PCR was expected to generate a product in transformed cells but not for the WT. A, Illustration of WT H. salinarum genotype. B, illustration of the mutant genotype after incorporation of plasmid pMG501. C and D, Agarose gel electrophoresis of PCR products using primers P1 and P2, and primers P3 and P2, respectively. Lane 1: MW markers (in bp), lane2: Chromosomal DNA of R1, lane 3: plasmid pMG501 (control), lanes 4–6- transformed colonies. E, Relative expression of OE1472F transcription in mutant StopOE1472F (OE1471F::pMG501) grown in synthetic medium without AroAA to OD600nm = 0.4 and 1.0. Results were obtained using RT-qPCR. See table S7 in file S1 for details of the primers used.

Figure 3