SCAR分子标记对水貂食毛症遗传病因的初步研究

Preliminary Studies on Genetic Etiology of Mink Trichophagia by SCAR

本研究利用序列特异性扩增(SCAR)技术对健康和食毛水貂进行遗传分析,旨在从分子水平探讨水貂食毛症的发病原因。首先从126个随机引物中筛选出4个重复性好的标记引物,通过引物A20和S139的扩增在两群体中找到差异标记(A20-1000,S139-400),对其进行克隆、测序,并根据测序结果设计2对SCAR引物,回到原样本群体中扩增验证。结果,SA20-757在食毛和健康水貂群体的分布频率分别为86.88%和27.41%,差异极显著(P<0.001);MS139-230在食毛和健康水貂群体的分布频率分别为94.21%和39.38%,差异极显著(P<0.001)。SA20-757与MS139-230联合分析表明,特异性条带在患病水貂群体的分布频率高达95.36%,均比单个SCAR特异引物的扩增比例要高。结果提示,SA20-757与MS139-230联合诊断分析,可以很好的区分健康与食毛水貂群体,可初步作为水貂食毛症早期诊断的分子生物学手段,为进一步研究水貂食毛症奠定基础。

In order to discuss the causes of mink trichophagia at molecular level,SCAR（sequenced characterized amplified region）technology was used to analyze genetic constitution of the healthy and trichophagia mink groups.Four polymorphic primers were screened from 126 random primers by RAPD.The different bands were amplified in healthy and sick mink groups.The specific bands were then cloned and sequenced,two SCAR primers were designed according to their sequence information by primer A20 and S139.Then PCR amplification was carried out in the healthy and trichophagia mink groups.The results showed that the frequency of SA20-757 in the two groups was 86.88% and 27.41%,respectively,and χ2 test of polymorphic fragments in the two groups showed significant difference（P〈0.001）;The frequency of MS139-230 in the two groups was 94.21% and 39.38%,respectively,and χ2 test of polymorphic fragments in the two groups showed significant difference（P〈0.001）.SA20-757 and MS139-230 associated analysis showed that the frequency of the specific bands in trichophagia mink group reached 95.36%,and it was higher than a single specific primer SCAR.These results indicate that the associated analysis of SA20-757 and MS139-230 can identify the healthy and trichophagia mink populations,and this molecular biological method can be used for the early diagnosis of trichophagia mink.