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. 2010 Oct 8;154(4):1672–1685. doi: 10.1104/pp.110.162990

Figure 1.

Figure 1.

Disruption of glutathione biosynthesis in Synechocystis 6803. A, Diagram of the glutathione biosynthetic pathway. B, The entire open reading frame of the gshA gene was replaced with a kanamycin resistance cassette (KmR) to generate the ΔgshA::KmR strain. C, Segregation of ΔgshA::KmR was tested by PCR using primers GshA5 and GshA6 shown in A. Lanes show wild-type genomic DNA (WT), pSL2083 (+), and ΔgshA::KmR genomic DNA. D, The gshB gene was replaced with a gentamicin resistance cassette (GmR). E, Segregation of ΔgshB::GmR was confirmed by PCR using primers GshB5, GshB6, and GshB7 shown in D. Lanes show wild-type genomic DNA (WT), pSL2085 (+), and ΔgshB::GmR genomic DNA. F, Growth of wild-type cells in the presence of the GshA inhibitor BSO. G, Cellular GSH concentration after 96 h of growth in the presence of BSO. Primer sequences used in cloning and segregation analysis are shown in Table I.