云斑尖塘鳢微卫星分子标记的筛选与检测

用磁珠富集法分离云斑尖塘鳢的微卫星序列,由所获得的1032个克隆中筛选出146个阳性克隆,经测序81个含有微卫星序列,52个为完美型,27个为非完美型,2个为复合型,其中43个微卫星序列重复次数在10以上。所获得的云斑尖塘鳢微卫星序列中除探针中使用的CA重复单元和GA重复单元外,还有TAC等其他类型的重复单元。设计合成38对微卫星引物,其中29对引物可稳定扩增出条带,使用这些引物对云斑尖塘鳢48个个体进行检测显示:观测杂合度平均值为0.63,期望杂合度平均值为0.43。29对引物中1对引物表现为单态,7对表现为高度多态,14对表现为中度多态,7对表现为低度多态,多态性较为丰富,说明本研究开发的绝大部分微卫星分子标记较为理想。

Isolation and Detection of Microsatellite Markers in Marbled Sleeper Oxyeleotris marmoratus

Microsatellite molecular markers were isolated in marbled sleeper Oxyeleotris marmoratus by magnetic-bead enrichment method,and 146 clones were sequenced from all the 1032 positive clones.Eighty-one sequences(55.5%) were shown to contain microsatellite in these clones,52 perfect type microsatellites,27 imperfect type microsatellites and 2 compound type microsatellites.Totally,43 sequences were found to contain more than 10 times microsatellite repeat units.These microsatellites contained not only CA and GA repeated unit,but a few other type unit(TAC) detected.Thirty-eight microsatellite primer pairs were designed and synthesized,in which 29 loci showed clear and good repeatability bands after testing.A population of 48 individuals was tested with the 29 loci.There were average observed heterozygosity(Ho) of 0.63 and expected heterozygosity(He) of 0.43.There were 7 highly polymorphic loci,fourteen moderate polymorphic loci revealed,seven low-grade polymorphic loci and one monomorphic loci in the population,indicating that most of the microsatellite loci from the fish are abundant polymorphology.