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利用抑制消减杂交初步构建犬小孢子菌在不同诱导底物表达的差异基因文库
  • ISSN号:1001-7089
  • 期刊名称:《中国皮肤性病学杂志》
  • 时间:0
  • 分类:R392.13[医药卫生—免疫学;医药卫生—基础医学] R379[医药卫生—病原生物学;医药卫生—基础医学]
  • 作者机构:[1]大连医科大学附属一院皮肤科,辽宁大连116011, [2]大连市友谊医院皮肤科工作,辽宁大连116031
  • 相关基金:国家自然科学基金(30640016)
中文摘要:

目的比较犬小孢子菌在儿童与成人头皮组织、儿童头皮组织与儿童光滑皮肤组织为诱导底物表达的差异基因,探讨犬小孢予菌蛋白酶在头癣中的发病机制。方法将犬小孢子菌菌株分别接种于含儿童光滑皮肤组织(driver1)、儿童头皮纽织(tester)和成人头皮组织(driver2)三种底物诱导培养基中培养,经抑制消减杂交技术(SSH)分别构建在tester与driver1,tester与driver2培养基中培养的犬小孢子菌菌株正反向差异基因文库,菌液PCR筛选鉴定,对阳性克隆进行测序分析。结果在tester与driver1的正向差异基因文库中初步筛选出1个差异表达基因与编码红色毛癣菌线粒体细胞色素b和NADH1-5具有高度同源性,另15个差异表达基因在真菌界均未知。结论应用SSH技术成功构建了不同底物诱导培养基犬小孢子菌表达的差异基因文库;这个与编码红色毛癣菌线粒体细胞色素b和NADH1-5具有高度同源性的差异表达基因,可能在犬小孢子菌所致头癣中起一定作用,进一步研究该基因的功能对研究头癣的发病机制具有重要意义。

英文摘要:

Objective To compare differential expressed cDNA genes of M. canis incubated on mineral medium with child's glabrous skin, scalp tissue or adult's scalp tissue by using SSH technique and to study the pathogenic mechanism of tinea capitis caused by microsporum canis. Methods Microsporum canis was cultivated on mineral medium with child's glabrous skin, scalp tissue and adult's scalp tissue as sole substrates . The forward and reverse cDNA library from microsporum canis incubated on mineral medium with tester and driverl, tester and driver2 were constructed by using SSH. The four subtractive library were screened firstly by PCR. The positive clones were sequenced and analysed. Results One of the twenty clones was high homologous with Trichophyton rubrum mitochondrial cytb gene and NADH1 to NADH5 genes. Fifteen of the twenty clones were unknown in fungus. Conclusion The cDNA libraries were constructed successfully from microsporum canis incubated in different proteases-induced medium successfully by using suppression subtractive hybridization (SSH). The screened clone that was high homologous with Trichophyton rubrum mitochondrial cytb gene and NADH1 to NADH5 genes may be on the role of tinea capitis. It is very meaningful to further study on this gene's function.

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期刊信息
  • 《中国皮肤性病学杂志》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国教育部
  • 主办单位:西安交通大学
  • 主编:彭振辉
  • 地址:陕西省西安市新城区皇城西路30号
  • 邮编:710004
  • 邮箱:CJDV_XG@126.com
  • 电话:029-87217312 87678312
  • 国际标准刊号:ISSN:1001-7089
  • 国内统一刊号:ISSN:61-1197/R
  • 邮发代号:52-17
  • 获奖情况:
  • 中国科技论文统计源期刊,中国生物医学类核心期刊,中国学术期刊综合评价数据库来源期刊,中国期刊方阵“双效”期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,美国剑桥科学文摘,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),中国北大核心期刊(2000版)
  • 被引量:25128