Isolation and characterization of 12 microsatellite loci in the noble scallop, Chlamys nobilis.

摘要

The isolation and characterization of 12 polymorphic microsatellite loci from the noble scallop (Chlamys nobilis) from a scallop farm in Shenzen, China, using polymerase chain reaction (PCR) were conducted. Primers were designed for each of 32 microsatellites with enough flanking sequences and more number of repeats. The number of alleles, heterozygosity, test of Hardy-Weinberg equilibrium (HWE) and linkage disequilibrium (LD) were analysed using GENEPOP 3.4. It was shown that of the 32 primer pairs tested, 10 did not generate expected PCR products, 9 produced multibands or complicated patterns, 1 yielded a monomorphic band over individuals and12 amplifed clear and specific products, showing polymorphism in the sample population tested in the study. Out of these 12 loci, 4, 5 and 3 were di-, tri- and tetra-nucleotide repeats, respectively. The average allele number of these polymorphic markers was 6.08 per locus, with a range of 3-12. Locus Cn102 was the least polymorphic (with 3 alleles) and Cn021 the most polymorphic (with 12 alleles). The values of observed heterozygosity varied from 0.22 to 0.93 and the expected ranged varied from 0.24 to 0.88, respectively. No significant linkage disequilibrium was detected for all the 12 polymorphic loci. This is the second set of microsatellite markers developed for this species so far. These microsatellite loci would be useful for stock study and genetic analysis of C. nobilis. More variable microsatellite loci are required in the future for studies such as pedigree analysis and stock management at a fine scale.