Structural Characterization and Molecular Cloning of Bioactive Peptides from the Skins of Amphibians Used in Traditional Medicine
Amphibian skins have been used for centuries as components of traditional medicines due to their possession of a wide spectrum of pharmacological effects．In recent years，analytical technologies have shown that they contain a plethora of active compounds，especially peptides．As amphibians are currently in global decline，it was the aim of our research to critically evaluate several methods oftissue／skin secretion preparation that would permit subsequent parallelpeptidome and transcriptome analyses，preferably without recourse to sacrifice of the donor specimen．cDNA libraries were constructed from both lyophilised venom and macerated air-dried skin using magnetic oligo—dTbead-captured P olyadenylated mRNA as templates．Those CDNAs encoding skin gra nular gland—derived pe ptideprecursors were amplified by a RACE PCR strategy．Products from different species of amphibian were purified，clonedand sequenced．Subsequent to prediction ofm ature P eptide amino a cid sequences from re spective cloned pr ecursorcDNAs，the presence of each peptide in the original preparation(skin secretion or dried skin)from each amphibian specieswas confirmed by a combination ofreverse—phase HPLC fractionation，MALDI-TOF mass spectrometry and automatedEdman degradation microsequencing．
peptides amphibian mass spectrometry molecular cloning
Mei Zhou Tianbao Chen Chris Shaw
Natural Drug Discovery Group，School of Pharmacy,Queen”s University Belfast，97 Lisbum Road，Belfast BT9 7BL，Northern Ireland，UK
国内会议
兰州
英文
402-403
2012-08-14（万方平台首次上网日期，不代表论文的发表时间）