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Similar amplifications of the ORF of CYP3A4 did not yield any splice variants (data not shown).
The hydropathy plot was not significantly altered by other variants (data not shown).
In contrast there was no decrease in the number of cell-associated bacteria when bacteria were treated with 0.25 µg/mL or 2.5 µg/mL of AbHV2BD linear versus AbHV2C linear, which does not bind OpaB variants (data not shown).
As had been determined previously for other proteins [14], [15], multiple tryptophan to phenylalanine replacements caused a serious reduction in stability of the variants (data not shown) and therefore variants containing multiple mutations were not analysed further in this study.
To explore whether the mutations made to the C-terminus protein sequence produced a change in secondary structure, circular dichroism was performed on all C-terminal variants (data not shown).
Assembly of the 5S rRNA gene confirmed three variants (data not shown); downstream of the 5S rRNA gene, all three variants differ from Methanoculleus marisnigri JR1, indicating a different genomic context for the rrn cluster.
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In support, we found the updated Roche GS Mapper software (version 2.3) correctly called the variant (data not shown).
When recombinantly expressing ADAR2R in HeLa and HEK293 cells, we did not detect any differences in general adenosine deaminase activity displayed by ADAR2R compared to the ADAR2a major splice variant (data not shown).
There was no variation among the wild type and its GFP variant (data not shown).
There was no relationship between any of the anti-CCP2 antibodies and the PTPN22 1858T variant (data not shown).
Of the 84 saturation mutagenesis variants chosen for FAME analysis, none were found to have more 16:1 in the medium than the M141T variant (data not shown).
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