毛囊蠕形螨与皮脂蠕形螨基因组DNA的RAPD分析和序列比对  

RAPD analysis and sequence alignment of genomic DNA of hair follicle mites Demodex folliculorum and D. brevis (Acari:Demodicidae)

作　　者：赵亚娥[1] 成慧[1] 

机构地区：[1]西安交通大学医学院免疫学与病原生物学系,西安710061

出　　处：《昆虫学报》2009年第11期1273-1279,共7页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30872199);陕西省自然科学基金项目(2006C247)

摘　　要：【目的】分析毛囊蠕形螨Demodex folliculorum(D.f.)和皮脂蠕形螨D.brevis(D.b.)基因组DNA的多态性,对相关条带进行测序分析。【方法】采用改良小昆虫DNA提取法提取两种人体蠕形螨基因组DNA,选择RAPD技术对其进行多态性分析,将相关条带分别与pMD18-T载体连接,克隆、测序后进行酶切鉴定和分析。【结果】毛囊蠕形螨共扩增15条带,皮脂蠕形螨共扩增12条带;两种蠕形螨既有共有条带,又有特异性条带;根据条带差异计算得到两种间的遗传距离为0.5556。毛囊蠕形螨约800bp处特异性条带测序结果显示,序列片段长度为855bp(GenBank登录号为FI277970);特异性引物扩增和酶切鉴定均为毛囊蠕形螨所特有。序列比对显示与阿糖胞苷DNA区域结合蛋白有46%的序列相似度。两种人体蠕形螨约300bp处共有条带序列分析显示,碱基序列均为341bp(GenBank登录号分别为D.f.FI520176;D.b.FI520175),在第84和第165位点有2个碱基不同,分别是A/G和C/T互换,同源性高达99.4%。但未发现有开放阅读框和相似度高的序列。【结论】序列片段为855bp的特异性条带为毛囊蠕形螨所特有;341bp碱基序列为毛囊蠕形螨和皮脂蠕形螨所共有,同源性高达99.4%。RAPD技术可用于两种人体蠕形螨基因组DNA的多态性分析和物种鉴定。[Objective] Analysis of genomic DNA polymorphism and the related sequence of Demodex folliculorum and D.brevis.[Methods] The genomic DNA of the human Demodex was extracted by using improved DNA extraction method of mini-insects.Random amplified polymorphic DNA(RAPD)was applied to analyze the polymorphism.The related bands were connected with pMD18-T vector,and cloned,sequenced,and identified and analyzed after enzyme digestion.[Results] There were 15 bands obtained in D.folliculorum and 12 in D.brevis.Some ...

关 键 词：毛囊蠕形螨 皮脂蠕形螨 RAPD分析 遗传距离 序列比对 物种鉴定 

分 类 号：Q96[生物学—昆虫学]