So far numerous antimicrobial peptides have been characterized from amphibians. In this work, a new family of antimicrobial peptides, named shuchin, was purified and characterized from skin secretions of the frog, Rana shuchinae that lives in freezing mountains. Totally two members of shuchin (shuchin 1 and 2) were identified with the amino acid sequence of NALSMPRNKCNRALMCFG and NALSSPRNKCDRASSCFG, respectively. cDNAs encoding shuchins were cloned from the skin cDNA library of R. shuchinae. The precursors of shuchin are composed of 62 amino acid residues including the conserved signal peptides, acidic propieces, and mature antimicrobial peptides. Synthetic shuchins showed strong and broad antimicrobial activities against Gram-positive bacteria (Staphylococcus aureus, and Bacillus cereus; MICs<12.5 microg/ml), Gram-negative bacteria (Escherichia coli, Bacillus dysenteriae, Pseudomonas aeruginosa; most MICs from 3.1 to 12.5 microg/ml), and yeast (Candida albicans; MICs of 6.25 microg/ml), but no hemolytic activity under the effective concentration, thereby provide more leading templates for designing novel anti-infection agents.