Fig. 2
Genomic maps of the engineered Synechocystis sp. PCC 6803 strains, KS and KAS. A Double homologous recombination occurred between the conserved sequences of sll1951 or S-layer gene on the recombinant pJSKm plasmid containing an antibiotic kanamycin resistant cassette (Kmr) and genomic DNA of WT or KA host strain, generating KS or KAS strain, respectively. Confirmations of engineered strains were performed by PCR analysis using selected pairs of specific primers (shown in Table 2). For (B) KS strain; Lane M: GeneRuler DNA ladder (Fermentus); Lane 1: Negative control using WT as template (a–c), a Lanes 2–3: clone numbers 1 to 2 using Km_F and Km_R primers, b Lanes 2–3: clone numbers 1 to 2 using Sll1951_F and Sll1951_R primers, and c Lanes 2–4: clone numbers 1 to 3 using Sll1951_UF and Sll1951_R primers, and only positive clones (numbers 2 and 3) were selected for next experiments. For (C) KAS strain. Lane M: GeneRuler DNA ladder (Fermentus); Lane 1: Negative control using KA as template (a–c), a Lanes 2–5: clone numbers 1 to 4 using Km_F and Km_R primers, b Lanes 2–5: clone numbers 1 to 4 using Sll1951_UF and Km_SR primers, and c Lane 2: clone number 1 using Sll1951_UF and Sll1951_R as the primers, and only positive clone number 1 was selected for next experimentsq4