Fig. 1
From: Enhancement of membrane vesicle production by disrupting the degP gene in Meiothermus ruber H328
Plasmid constructions (A) and homologous recombination (B) of M. ruber H328. ( A) A hybrid plasmid pUC119-∆degP-htk for homologous recombination was constructed by using the vector plasmid pUC119 and three DNA fragments (1.0 kbp each) including the 5’-flanking and 3’-flanking regions of the degP gene (mrH_0331) in the chromosomal DNA of strain H328 and a thermophilic kanamycin-resistant gene (htk) from the plasmid pUC18-htk, respectively. DNA fragments were obtained by PCR by using a pair of primers listed in Additional file 1: Table S1 in the reaction condition as described in the Materials and methods section and their digestion and ligation followed the procedure recommended by the vendor. (B) Homologous recombination was carried out as described in the Materials and methods section, and the knockout of the degP gene was confirmed by PCR using a pair of primers (degP/F1-Hind and htk/Rv-Pst or htk/Fw-Pst and degP/R4-Xba)