With respect to the reference genome, β1 strains had between 688 and 9,828 SNPs and β2 strains had between 17,355 and 21,071 SNPs (Figure 2). In the β1 strains the number of SNP-dense
regions was low, whereas there were many more SNPs in the β2 strains due to their greater phylogenetic difference from the reference. The single ψ strain had 32,828 SNPs (not shown in Figure 2). Although the β2 strains and the ψ strain had a broadly similar number of SNPs, they were clustered in patterns that were distinct between the groups, a finding consistent with regions of high SNP density likely representing distinct recombination events. We hypothesised that “blocks” of NVP-LDE225 DNA sequence with a high frequency of SNPs, separated by regions of the genome with low SNP density, could each represent an individual transformation event (Figure 2). To investigate this, we analysed two strains (RM7578 and RM7122) that have the same multi-locus sequence type. RM7578, the strain most closely related to the reference strain 10810, has five blocks of SNPs. For this analysis, blocks were defined as contiguous regions containing at least 30 SNPs, with each SNP separated by mTOR inhibitor no more than 300 bp. 91%
of 694 SNPs between strains RM7578 and 10810 were found within these five blocks, amounting in total to 23.5 kbp of DNA, or 1.2% of the genome. Strain RM7122 had 15 blocks of SNPs when compared to strain 10810, equivalent to 2.4% of the genome. In the β1 strains, the size of these blocks ranged from less than 0.5 to more than 25 kbp, with a median size of 4.8 kbp (Figure 3), findings within the range recently reported experimentally for H. influenzae strains . We concluded that the blocks of SNPs identified between the closely related Hib strains represented recombination events, click here resulting in allelic exchanges that could delete or insert novel DNA, including whole genes. Figure 3 Size of SNP blocks found in the β1 group of Hib
strains. This histogram represents the frequency of different sizes of SNP blocks (as defined in the text) in the genomes of β1 H. influenzae type b strains. Inserted or deleted regions of DNA in Hib strains, relative to the genome sequence of reference strain 10810, were identified by BLASTN searches and the ACT genome browser. For a closely related strain, DC800, an example of insertion of a novel block of SNPs, mediated through transfer of DNA from an unknown donor, was identified. This inserted DNA included a putative gene flanked by regions of significant similarity. As a further example, comparison between two more divergent genomes (RM7060 and 10810) revealed at least 16 regions of DNA, each over 500 bp in length, that were present in one strain but not the other (Table 2). These regions constitute over 17.