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Figure 4 | BMC Microbiology

Figure 4

From: Biochemical and functional characterization of SpdA, a 2′, 3′cyclic nucleotide phosphodiesterase from Sinorhizobium meliloti

Figure 4

Purified Clr binds 3′, 5′cAMP and 2′, 3′cAMP nucleotides in vitro. (A) Clr-GST purification on a glutathione sepharose column. 1: Molecular weight markers, 2: Bacterial sonication pellet, 3: Sonication supernatant, 4: Column flowthrough, 5: Column wash, 6: Purified Clr-GST, 7: Clr-GST concentrated on centricon CO10000. (B,C) Clr affinity chromatography on a 3′, 5′cAMP-Agarose column (Sigma) and fraction analysis by SDS-PAGE (4-12%). 1: Molecular weight markers, 2: Free Clr (load), 3 : Flowthrough, 4-10: column wash, 11: eluted fraction by either 30 mM 3′, 5′cAMP (B) or 30 mM 2′, 3′cAMP (C).

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