Fig. 1

Construction of expression vector containing BuLIF. A. Vector map of pET22b(+) pelB/BuLIF constructed via Lasergene software. BuLIF encoding 540 bp without signal sequence was in cooperated into pET22b(+) vector and named as pET22b(+) pelB/BuLIF. The constructed expression vector encodes an N-terminal hexahistidine (His6) fused to the BuLIF (His6-BuLIF). This construct was transformed into Top 10F′competent E. coli cells. B The presence of BuLIF gene was confirmed by colony PCR and double restriction enzymes digestion analysis using NCoΙ and Xho Ι. Bands of 540 bp from different recombinant plasmids were observed in agarose gel. C Upon restriction digestion, two products of 540 bp BuLIF and 5493 bp pET22b(+) vector were released from recombinant plasmid confirming the presence of BuLIF in the expression vector. Uncut plasmid was run alongside the digested recombinant plasmid