Extended Data Fig. 8: Recombination between the genes BT1040, BT1042, and BT1046 and effect of BT1033-52 locus.
Recombination between the genes BT1040, BT1042, and BT1046 and effect of BT1033-52 locus. a, Pfam domain schematics of the amino acid sequences of these three genes highlighting that BT1040 and BT1046, as originally assembled in the B. thetaiotaomicron genome sequence, lack additional N-terminal sequences that are present on BT1042. b, Sequencing of the 8 PCR amplicons schematized in Fig. 5d. Amplicons 1, 5 and 8 represent the original genome architecture, while the others represent inferred recombination events that are validated here by sequencing. The 5′ and 3′ ends of the BT1042, BT1040 and BT1046 genes are color-coded to assist in following their connectivity changes after recombination. A series of single-nucleotide polymorphisms (SNPs) present in BT1042, downstream of the proposed recombination site, are highlighted in yellow. The transfer of these SNPs to a fragment containing the 5′ end of BT1040 (Amplicon 4) was used to narrow the recombination region to the 7 nucleotide sequence highlighted in red. Additional SNPs that are specific to the regions upstream of this recombination site are shown in white text for each sequence. Susceptibility of acapsular B. thetaiotaomicron to ARB25 without the BT1022-52 locus is not affected. c, Ten-fold serial dilutions of ARB25 were spotted onto lawns of B. thetaiotaomicron ∆cps (n = 5) and B. thetaiotaomicron ∆cps ∆BT1033-52 (n = 5). Each of the 5 biological replicates contained 3 technical replicates from independent clones and all 15 replicates are shown individually. Plaquing efficiency was determined by normalizing plaque counts on B. thetaiotaomicron ∆cps ∆BT1033-52 relative to plaque counts on B. thetaiotaomicron ∆cps for each replicate. Statistical significance was determined using the 2-tailed Mann-Whitney test and bars represent the geometric mean.
