FIG 1 .

Construction of CA_P0037::int mutants. (A) DNA configuration of mutant with a group II intron inserted at position 189/190 on the sense strand of CA_P0037. (B) PCR on genomic DNA of CA_P0037::int mutant (MT) and wild-type (WT) strains with different pairs of primers (intron probe-F and CAP0037-Fb primers and CAP0037-Rb and CAP0037-Fb primers) to confirm intron insertion and primers LtrA 5D and LtrA 3R to demonstrate the loss of pCUI1-cap37(189s). PCR was performed on genomic DNA of complemented mutant (MTC) strain with primers pthla-MF and cap37-sfo1-R to demonstrate the presence of the pSOS95-Cap0037 plasmid. (C) Detection of the Cap0037 protein in the wild type (WT), CA_P0037::int mutant (MT), and CA_P0037::int complemented mutant (MTC). The Western blot was generated using polyclonal antibodies raised against purified His-tagged Cap0037. The positions of molecular mass markers (in kilodaltons) are shown to the left of the Marker blot. (D) Southern blot to demonstrate single intron insertion in the CA_P0037::int mutant. The intron probe was DIG labeled and hybridized to EcoRI-digested genomic DNA of the CA_P0037::int mutant (lane 1) and to the AgeI-SbfI-digested pCUI1-cap37(189s) plasmid as a positive control (lane 2) with expected sizes of 4,388 bp and 1,718 bp, respectively.