Isolation and Sequence Characterization of Microsatellite loci in Neosalanx taihuensis
Neosalanx taihuensis is naturally distributed in the middle and lower reaches of the Yangtze River and its connected lakes.It is one of the most important economic fishes in these waters and has been widely and success-fully introduced to other fresh water lakes and reservoirs.Studies on the species mainly focused on stock enhance-ment,but there is a lack of research on its genetics which could be important for resource management.To better understand the genetic diversity and genetic structure of Neosalanx taihuensis,microsatellite DNA markers were de-veloped for N.taihuensis using the magnetic bead technique.(AC)12 ,(AAC)10 ,(AAG)10 and (GATA)8 oligo-nucleotide probes were used to screen the microsatellite sequences in six N.taihuensis specimens collected from Fuxian Lake and Taihu Lake in March,2012.A total of 490 microsatellite sequences were identified with a total screening efficiency of 78.09%.A total of 725 microsatellite loci were detected,with 592 perfect repeat motifs ac-counting for 81.66%,82 compound repeat motifs accounting for 11.31%,and 51 imperfect repeat motifs account-ing for 7.03%.Among them,there were 390 di-nucleotide repeat loci,284 tri-nucleotide repeat loci,47 tetra-nu-cleotide repeat loci,1 penta-nucleotide repeat locus and 3 hexa-nucleotide repeat loci,contributing,respectively, 53.79%,39.17%,6.48%,0.14% and 0.42% to the total microsatellite loci.The number of microsatellite re-peats enriched by probe (AC)12 vanged mostly from 14 to 26 but with the highest number of 44.The number of mi-crosatellites repeats enriched by probes (AAC)10 or (AAG)10 ranged mostly from 8 to 20 with the highest number of 42.While the microsatellites repeats enriched by probes (GATA)8 ranged mostly from 6 to 15 with the highest number of 32.The hybridization efficiency of probes (AC)12 ,(AAC)10 ,(AAG)10 and (GATA)8 was 85.19%, 60.19%,5.16% and 10.64%,respectively.PCR amplification was then conducted on 24 samples of N.taihuen-sis collected from Fuxian Lake using the primer design obtained from the di-nucleotide repeat loci.The electro-phoretic pattern of PCR products showed high polymorphism.The tri-nucleotide repeat loci and tetra-nucleotide re-peat loci obtained will be useful for developing long repeat microsatellite loci for N.taihuensis.
Neosalanx taihuensismicrosatelliteenrichment by magnetic beads
国家科技期刊平台
NSTL
万方数据
维普
