Brevinins　are　peptides　of　24　amino　acid　residues,　originally　isolated　from　the　skin　of　the　Oriental　frog,　Rana　brevipoda　porsa,　by　nature　of　their　microbicidal　activity　against　a　wide　range　of　Gram-positive　and　Gram-negative　bacteria　and　against　strains　of　pathogenic　fungi.　cDNA　libraries　were　constructed　from　lyophilized　skin　secretion　of　three,　unstudied　species　of　Chinese　frog,　Odorrana　schmackeri,　Odorrana　versabilis　and　Pelophylax　plancyi　fukienensis,　using　our　recently　developed　technique.　In　this　report,　we　describe　the　＂shotgun＂　cloning　of　novel　brevinins　by　means　of　3＇-RACE,　using　a　＂universal＂　degenerate　primer　directed　towards　a　highly　conserved　nucleic　acid　sequence　domain　within　the　5＇-untranslated　region　of　previously　characterized　frog　skin　peptide　cDNAs.　Novel　brevinins,　deduced　from　cloned　cDNA　open-reading　frames,　were　subsequently　identified　as　mature　peptides　in　the　same　samples　of　respective　species　skin　secretions.　Bioinformatic　analysis　of　both　prepro-brevinin　nucleic　acid　sequences　and　translated　open-reading　frame　amino　acid　sequences　revealed　a　highly　conserved　signal　peptide　domain　and　a　hypervariable　anti-microbial　peptide-encoding　domain.　The　experimental　approach　described　here　can　thus　rapidly　provide　robust　structural　data　on　skin　anti-microbial　peptides　without　harming　the　donor　amphibians.　(c)　2005　Elsevier　Inc.　All　rights　reserved.