Fig. 11
a Double digestion of recombinant plasmid pET28a-PspA1-5c+p with NcoI and XhoI restriction enzymes. Lane1. 1kB DNA Ladder marker, 2. Undigested plasmid, 3. Double digestion of the plasmid with NcoI and XhoI, resulting in two bands of the PspA1-5c+p (~ 1826 bp) and pET28a (~ 5639 bp). 3. Monodigestion of plasmid with NcoI. b Colony PCR on transformed E. coli BL21 colonies with universal T7 primers. Lane1. 1kB DNA Ladder marker, lanes 2 to 6. Positive colonies with a band size of the PspA1-5c+p (~ 1826 bp). c Purification of recombinant PspA1-5c+p protein by Ni–NTA chromatography. d Western blot analysis of the purified recombinant PspA1-5c+p