CAJ | 학술논문

利用FIASCO (Fast Isolation by AFLP Sequences Containing repeats)技术建立拟穴青蟹Scylla paramamosain 基因组文库,并与生物素标记的(CA)15寡核苷酸探针杂交,联合磁珠富集法构建拟穴青蟹微卫星富集文库。测序194个阳性菌落,分析其中的150条序列,结果表明：两碱基重复类型占90%以上,其中重复拷贝数在30以上的占27.45%；含微卫星座位189个,其中完美型146个、非完美型28个和复合型15个。设计125对引物扩增一个拟穴青蟹野生群体(含20个个体),其中的19对引物能稳定扩增且片段大小基本符合理论长度。遗传变异分析表明,17个位点表现出高度多态性,16个位点显著偏离Hardy-Weinberg平衡(P<0.05),4组两两位点间存在连锁不平衡现象(P<0.0026,经Bonferroni法校正),7个微卫星位点可能存在无效等位基因。若排除混合微卫星位点的引物对以及扩增位点 PIC (polymor-phism information content)值在0.5以下的引物对,则13对引物能用于拟穴青蟹群体遗传学等研究。
이용FIASCO (Fast Isolation by AFLP Sequences Containing repeats)기술건립의혈청해Scylla paramamosain 기인조문고,병여생물소표기적(CA)15과핵감산탐침잡교,연합자주부집법구건의혈청해미위성부집문고。측서194개양성균락,분석기중적150조서렬,결과표명：량감기중복류형점90%이상,기중중복고패수재30이상적점27.45%；함미위성좌위189개,기중완미형146개、비완미형28개화복합형15개。설계125대인물확증일개의혈청해야생군체(함20개개체),기중적19대인물능은정확증차편단대소기본부합이론장도。유전변이분석표명,17개위점표현출고도다태성,16개위점현저편리Hardy-Weinberg평형(P<0.05),4조량량위점간존재련쇄불평형현상(P<0.0026,경Bonferroni법교정),7개미위성위점가능존재무효등위기인。약배제혼합미위성위점적인물대이급확증위점 PIC (polymor-phism information content)치재0.5이하적인물대,칙13대인물능용우의혈청해군체유전학등연구。
Genomic library of Scylla paramamosain (mud crab) was first constructed by FIASCO (Fast Isolation by AFLP Sequences Containing repeats) technique and then was hybridized to (CA)15 biotin-labeled probe. The magnetic bead enrichment method was employed to capture the target fragments for microsatellite DNA enrichment library. A total of 194 positive bacterial colonies were sequenced, among of which 150 sequences were investigated. As a result, dinucleotide repeat preponderated over 90%, whose repeat copy more than 30 times accounted for 27.54%. 189 microsatellite sites were found, containing 146 perfect type, 28 imperfect type, and 15 compound type. The polymorphisms of 125 primer pairs were estimated through amplifying a solo wild population of Scylla paramamosain (containing 20 individuals). 19 loci showed stable amplification, whose products conformed to theory size basically. Genetic varaiability analysis suggested 17 highly polymorphic loci. 16 loci were deviated from Hardy-Weinberg equilibrium significantly (P<0.05). Significant linkage disequilibrium was found in four pairwise loci (P<0.0026, corrected by Bonferroni method) while null alleles were detected at seven loci. Regardless of the loci of microsatellite mixture (containing two microsatellite sites or over) plus some loci with PIC value lower than 0.5, 13 primer pairs proved to be useful for the population genetic analysis.