The unique cluster contained 49 ORFs, out of which 30 were hypothetical Epacadostat chemical structure proteins dispersed throughout the cluster. Conserved domain analysis of these hypothetical proteins showed that many of these had domains of phage-related proteins such as AraC-type DNA-binding domain containing protein (VCD_003673), TraW (VCD_003693), PglZ (VCD_003717). There were also hypothetical proteins having potential domains of uracil-DNA glycosylase (VCD_003689), PLDc (VCD_003699), GP4d helicase (VCD_003701), type II restriction enzyme (VCD_003718), putative inner membrane protein (VCD_003722), MFS (VCD_003735), HATPase_c (VCD_003751).

Apart from the hypothetical proteins there were integron integrase (VCD_003670), transposase at VCD_003728 and VCD_003743 and a IS phage Tn transposon-related protein

at VCD_003742. There were phage-related proteins such as TraF, TraD and TraI. Along with these, there were other biosynthetic, regulatory and transferase-like proteins as well (Table S1). Analysis of unique horizontal gene transfer (HGT) regions as shown in Fig. 2b revealed that the Classical strain had the highest percentage of unique ORFs in the predicted GIs (7%), whereas V. cholerae El Tor N16961 had only 1% of unique ORFs in the PD0332991 order predicted GIs. Interestingly, V. cholerae MJ1236, which is regarded as a hybrid strain between Classical MYO10 and El Tor, had a high percentage of unique HGTs (5%). This led us to believe that it had undergone incorporation of GIs not only from Classical and El Tor

but also from other sources as well. It appears that the V. cholerae genomes had undergone several genetic modifications over time, explaining their diversity in pathogenicity and pandemicity. The genomes had been very dynamic with substantial changes through mutation, recombination, acquisition of genes in islands and acquisition of cassettes in the major integron (Karaolis et al., 1999). The present study showed that there were regions that were shared by all the three strains under study; however, each of the strains revealed regions that were unique to them. Our study revealed that V. cholerae MJ1236 shared distinct GIs with the V. cholerae Classical strain O395 that were not present in the V. cholerae El Tor strain and vice versa. The study also indicated that a greater percentage of GIs of V. cholerae MJ1236 were shared with V. cholerae Classical strain O395 than with V. cholerae El Tor strain. Even though V. cholerae MJ1236 had a high percentage of GIs shared with either of the other two strains under study, it was interesting to note that distinct sets of GIs were present in the chromosomes of V. cholerae MJ1236 that were unique to it. Vibrio cholerae MJ1236 revealed a section of a GI containing a large cluster of ORFs that was not shared by the other two strains.