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The venom of Lycosoidea spiders is a complex multicomponent mixture of neurotoxic pep-tides(main components)and antimicrobial peptides(AMPs)as minor components.In this study,we described the high-throughput identification and analysis of AMPs from Lycosa sinensis venom(named LS-AMPs)using a combination strategy that includes the following three differ-ent analysis approaches:(i)peptidomic analysis,namely reversed-phase high-performance liquid chromatography(RP-HPLC)separation plus top-down sequencing by matrix-assisted laser desorp-tion/ionization time-of-flight mass spectrometry(MS);(ii)transcriptomic analysis,namely cDNA library construction plus DNA sequencing;(iii)bioinformatic analysis,namely analysis and predic-tion for molecular characters of LS-AMPs by the online biology databases.In total,52 sequences of AMPs were identified from L.sinensis venom,and all AMPs can be categorized into eight different families according to phylogenetic analysis and sequence identity.This is the largest number of AMPs identified from a spider species so far.In the present study,we demonstrated molecular char-acteristics,such as complex precursor,N-and/or C-terminally truncated analogs,and C-terminal amidation of LS-AMPs from L.sinensis venom.This is a preliminary investigation on the molecu-lar diversification of venom-derived AMPs from the wolf spider species(family Lycosidae),and a detailed investigation on the functional diversity of LS-AMPs will be preformed in the future.