Rapid isolation and characterization of polymorphic microsatellite loci in the mud crab, Scylla paramamosain (Portunidae).

Scylla paramamosain is a widespread and commercially important species of coastal marine crab. We identified 13 polymorphic microsatellite loci from a genome library constructed with 5'-anchored PCR method. Thirty-two S. paramamosain from the East China Sea were used to analyze the characteristics of these loci. The number of alleles per locus ranged from 3 to 8, with a mean of 5.923. Observed and expected heterozygosities ranged from 0.500 to 0.875 and from 0.500 to 0.859, respectively. Eleven of the 13 loci were highly polymorphic (polymorphic information content >0.5). All of the 13 novel loci were in Hardy-Weinberg equilibrium after Bonferroni's correction (P < 0.0038). There was no null allele, stuttering errors or evidence of allelic dropout in any of the loci analyzed by MICRO-CHECKER. According to pairwise tests, no significant linkage disequilibrium was found among the 13 loci (P < 0.0038, adjusted value). These novel developed microsatellites will be useful for studies of genetic variation, population structure, conservation genetics, and molecular-assisted selective breeding of S. paramamosain.

Homology cloning, sequence characterization, and expression analysis of cDNA encoding electron transfer flavoprotein beta polypeptide in mud crab (Scylla paramamosain).

Electron transfer flavoproteins (ETFs) are αß-heterodimers found in eukaryotic mitochondria and bacteria. Herein we report a full-length complementary DNA of a mud crab (Scylla paramamosain) ETF ß subunit (Scpa-ETFB) isolated with a homology cloning strategy. The complete complementary DNA of the Scpa-ETFB contains a 17-nt 5'-untranslated region, a 765-nt open reading frame encoding 254 amino acids, and a 248-nt 3'-untranslated region. The high identity of Scpa-ETFB with ETFB in other organisms indicated that Scpa-ETFB is a new member of the ETFB family. Although the conserved motif associated with flavin adenine dinucleotide binding is absent in Scpa-ETFB, the signature sequences of the ETF superfamily were identified. Using reverse transcriptase polymerase chain reaction, we detected the messenger RNA transcript of Scpa-ETFB in high levels in the tissues of the hepatopancreas, ovary, heart, and muscle. Phylogenetic analysis showed that Scpa-ETFB is most closely related to the ETFB genes of Caligus rogercresseyi and Lepeophtheirus salmonis. These results provided basic information for elucidating the molecular mechanism of energy production in the mud crab.