The mudskipper, Boleophthalmus pectinirostris (Perciformes, Gobiidae), is an amphibious gobioid fish and a good species for marine culture in China. In this paper, 16 polymorphic microsatellite loci were isolated and characterized. The markers were tested on 60 individuals collected from Beihai (BH) and Lianyungang (LYG) coast in China. The number of alleles (N A ) in these markers ranged from 5 to 11. The polymorphism information content varied from 0.5946 to 0.8567 in the two populations. The observed (H O ) and expected (H E ) heterozygosity ranged from 0.5333 to 0.9000 in BH and from 0.5333 to 0.8729 in LYG, respectively. No significant linkage disequilibrium was detected at any pair of loci (P > 0.01). These microsatellite loci will be useful for population genetic studies of B. pectinirostris.