Fig. 2

Analysis of the MPER insert stability over the passages. (A) PCR analysis for the presence of MPER insertion in the bacterial genome of the EcN. Higher (compared to EcN) molecular weight bands confirm the presence of the insert in the genome of EcN-MPER bacteria. (B) Pairwise sequence alignment of the sequenced MPER insert in passages 1 and 30 of the recombinant EcN-MPER. A sequence similarity of 100% was obtained through analysis with the Needle (EMBOSS) Global Alignment Tool, which connotes an absence of mutation. (C) Western blot to evaluate the expression of the MPER in EcN-MPER and non-recombinant EcN. The whole cell lysate (lysate), soluble fraction in the supernatant (supern.) and insoluble cell debris (pellet) of passages 1 and 30 (P1 and P30) were analysed after probing with 2F5 antibodies. The expected size of recombinant OmpF-MPER is 43 kDa