The protein sequence similarity between two targets is calculated by sequence alignment and is defined as S t seq (t u, t v ) = align (t u, t v ) / align (t u, t u ) align (t v, t v ), where align(t u, t v ) is the Smith-Waterman alignment score [ 15] between t u and t v.
The homology of VGSCs was calculated by aligning sequences using ClustalW [24] followed by calculation of sequence identity using GeneDoc [25].
The distance matrix calculated by performaing sequence alignment between every pair of sequences within an input metagenome sample.
The minimum interspecific distances were calculated by comparing sequences of the species of interest to sequences of all remaining species in the dataset.
Specificity is calculated by dividing sequence counts within the targeted region by total sequence counts mapable to the human genome uniquely for each run.
Ct value was calculated by the Sequence Detection System software (Applied Biosystems).
Theoretical molecular weight and p I of characterized proteins were calculated by processing sequence entries at.
Average ratios were calculated by normalizing sequence coverage at T3 (65') and T4 (70') with T0 coverage.
Amino acid coverage was calculated by mapping sequences back to an assembled proteome.
Divergence was calculated by comparing sequence pairs in relation to the phylogeny reconstructed by MegAlign.> Additional file 1: Gel quantification analysis of all bands by ImageJ.
