Genetic Confirmation of the Role of Sulfopyruvate Decarboxylase in Coenzyme M Biosynthesis in Methanococcus maripaludis
Figure 2
Characterization of the comE::Tn5 mutant strain S201. (a) Genetic maps of the gene comE in the wild type strain and the comE::Tn5 mutant strain S201. Numbers indicate the MMP identification, and the black arrows indicate primers used for PCR amplification. (b) Genotypic characterization of the comE::Tn5 mutant strain 201 by PCR amplification. Lane 1, standard 1 kb ladder (New England Biolab); Lanes 2 and 3, PCR amplifications of comE using primers comEF (Af) and comER (Ar) for genomic DNA of the comE::Tn5 mutant strain S201 and wild type strain S2, respectively; Lanes 4 and 5, PCR amplifications using a primer from the end of the transposon (KAN-2RP-1out2; Br) and a primer that binds upstream of the gene comE (Bf) for the same DNAs. (c) Growth of the wild type and comE::Tn5 mutant strain 201 in minimal medium + acetate (McNA) and McNA supplemented with coenzyme M after three passages. Grey, comE::Tn5 McNA; green, comE::Tn5 McNA + CoM; red, S2 McNA; blue, S2 McNA + CoM. Representative error bars indicate the standard deviation of three independent cultures. The inoculum was ~1 × 104 cells.
