The　defensive　skin　secretions　of　amphibians　are　a　rich　source　of　bioactive　peptides.　Here　we　describe　a　rapid　technique　for　skin　granular　gland　transcriptome　cloning　from　a　surrogate　tissue-the　secretion　itself.　cDNA　libraries　were　constructed　from　lyophilized　skin　secretion　from　each　of　the　Chinese　frogs　(Rana　schmackeri,　Rana　versabilis,　and　Rana　plancyi　fukienensis)　using　magnetic　oligo(dT)　bead-captured　polyadenylated　mRNA　as　templates.　Specific　esculentin　cDNAs　were　amplified　by　3＇-RACE　using　a　degenerate　primer　designed　for　a　consensus　nucleotide　sequence　in　the　5＇　untranslated　region　of　previously　characterized　ranid　frog　peptide　cDNAs.　The　cloned　cDNAs　were　found　to　encode　the　antimicrobial　peptides　esculentins　1　and　2　from　each　of　the　species　examined.　The　presence　of　predicted　peptide　structures　in　skin　secretions　was　confirmed　by　MALDI-TOF　mass　spectrometry　and　automated　Edman　degradation.　This　experimental　approach　can　thus　rapidly　expedite　parallel　transcriptome　and　peptidome　analysis　of　amphibian　granular　gland　secretions　without　harming　or　sacrificing　donor　animals.　(c)　2005　Elsevier　Inc.　All　rights　reserved.