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Figure 2 | Journal of Biological Engineering

Figure 2

From: Inactivation of uptake hydrogenase leads to enhanced and sustained hydrogen production with high nitrogenase activity under high light exposure in the cyanobacterium Anabaena siamensis TISTR 8012

Figure 2

Engineering of a ∆hupS strain of Anabaena siamensis TISTR 8012. (A) Physical map of hupSL in wild type A. siamensis TISTR 8012 and an engineered strain lacking a functional uptake hydrogenase (∆hupS) created by interrupting hupS with neomycin antibiotic resistant cassette. (B) Confirmation of complete segregation of ∆hupS was performed by using colony PCRs and analyzed by 0.8% agarose gel electrophoresis. Primer pairs specific to hupS was used; Lane M: GeneRulerTM DNA ladder (Fermentas), Lane P1: Positive control (PCR product using genomic DNA of wild type as template), Lanes 1–3: PCR products of recombinant colonies cultured in BG11 medium in which the addition of antibiotic for 2 weeks did not show complete segregation, Lanes 4–6: PCR products of recombinant colonies cultured in BG11medium in which the addition of antibiotic for 4 weeks did show complete segregation, Lane P2: Positive control (PCR product using pRLhupSNm plasmid as template), Lane N: Negative control using H2O as template.

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