目的 剑尾鱼是由原良种委员会审定并由农业部公布的水生实验动物,在遗传学研究、水环境污染监测、细菌性疾病研究方面显示出较好的应用前景.为了对剑尾鱼选育系进行种质资源监测、区分选育系与非选育以及鉴定近交系纯度,本研究采用微卫星DNA进行水生实验动物剑尾鱼的指纹图谱构建.方法 根据相关报道设计合成了50对微卫星引物,对几个剑尾鱼品系的种质资源进行检测,筛选品系间差异性引物;确立剑尾鱼核心引物,用EXCEL散点图绘制DNA指纹图谱模式图;并将数字化指纹数据输入珠江水产研究所鱼类种质鉴定软件V1.0,形成剑尾鱼标准化指纹图谱鉴定数据库.结果 共获得剑尾鱼品系间特异标记5个可用于剑尾鱼近交系鉴定,确立46个微卫星标记为核心引物,构建剑尾鱼选育系RR-B系、RW-H系和非选育的野生品种的DNA指纹图谱.结论 本研究筛选出的微卫星标记与构建的指纹图谱,可用于剑尾鱼3个品种间的品种鉴定、纯度检测及遗传监测.%Objective Swrodtail, Xiphophorus hellerii was an aquatic experimental animal authorized by the original thoroughbred committee and published by the ministry of agriculture. It shows a good prospect in genetic research experiment, water environment pollution monitoring and bacterial disease research. In order to monitoring the germplasm resources of swordtail, distinguish the inbred lines and wild swordtail and identify the purity of inbred lines, microsatellite DNA fingerprint was used in this study. Methods 50 pairs of microsatellite primers were synthesized according to related reports. Among which, train-special primers used to identify the inbred lines of swordtail and core primers used to draw the DNA fingerprinting schema graph were screened. Then we constructed the DNA fingerprint using scatter diagram method with EXCEL 2003 software. Digital fingerprint barcode data had input software of PRIFRI V1.0, which had formed a standardization fingerprint identification database of swordtail. Results We together received five pairs of strain-special microsatellite primers and 46 core primers. The DNA fingerprinting of RR-B, RW-H and wild swordtail were constructed.And a fingerprint identification database of swordtail was formed. Conclusion Using these microsatellite markers and DNA fingerprinting, we can identify varieties, purity and genetic monitoring of RR-B, RW-H and wild swordtail.
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