水貂自咬症遗传病因的SCAR分子标记分析

Genetic Etiology Study of Mink Self-biting Disease by SCAR

以自咬和健康水貂作为研究对象,利用序列特异性扩增(SCAR)技术对其进行遗传分析,从分子水平探讨水貂自咬症的发病原因。首先从100个随机引物中筛选出10个重复性好的标记引物,对94只水貂群体进行随机扩增DNA(RAPD)标记检测。挑选出在健康组与患病组差异明显的A8引物,其仅可以在患病水貂组中扩增出500 bp左右的DNA片段,而在健康组中没有此特异片段。将该片段克隆、测序,根据测序结果设计特异PCR引物,转化为SCAR标记,回到原样本群体中扩增,验证。结果发现,MA8-500特异性条带在患病水貂群体的分布频率高达86.4%(38/44)以上,而在健康水貂群体中的分布频率仅为4.0%(2/50),因此可将其初步作为区分健康和患病水貂群体的分子遗传标记,为进一步研究水貂自咬症奠定基础。

In order to discuss the causes of self-biting disease at molecular level,SCAR（sequenced characterized amplified region）technology was used to analyze genetic constitution of the healthy mink and self-biting mink groups.RAPD（random amplification polymorphism DNA） technique was firstly adopted based on the reproducible 10 polymorphism primers screened from 100 random primers in the 94 mink groups.With the primer A8,different band was amplified between healthy and sick mink groups.The amplified 500 bp DNA fragment only appeared in the self-biting mink groups.The specific band was then cloned and sequenced,two primers were designed according to their sequence information to transform into SCAR marker.Then PCR amplification was carried out in the health mink groups and self-biting mink groups.The results showed that the frequency of MA8-500 specific band in self-biting mink groups reached 86.4%（38/44）,but only 4.0%（2/50） in health mink groups.The high frequency of self-biting mink groups can be served as molecular genetic label to distinguish preliminarily from healthy and self-biting groups,which lay a foundation for further study of mink self-biting disease.