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目的 进一步了解裴氏着色霉的基因学特征 ,采用溴苯提取法提取基因组DNA。方法 以随机扩增DNA多态性法对来源于 7个国家的 3 2株裴氏着色霉 (日本 15株 ,中国 3株 ,委内瑞拉 4株 ,中美洲 4株 ,哥伦比亚 2株 ,墨西哥 2株 ,巴西 1株 ,泰国 1株 )的DNA指纹图谱作了分析。结果 ①共选用 10个随机引物进行扩增 ,筛选出 3个具有较好DNA扩增片段的引物 ,即引物 1:5’ GAGCC CTCCA 3’ ,引物 7:5’ CGGCCCCTGT 3’ ,引物 8:5’ AGGTCACTGA 3’。② 3 2株裴氏着色霉的DNA带型不完全相同 ,具有一定的遗传变异性。③来源于同一国家的菌株具有较高的遗传相似性。④来源于日本的 15株菌株采用引物 8扩增后呈现两种DNA带型 ,I型 (有 3条主带 ) ,Ⅱ型 (有两条主带 ) ,其余国家菌株由于数量较少 ,DNA分型不明显。⑤同一引物扩增 ,90 %以上菌株可获共同的DNA片段。结论 随机扩增DNA多态性研究发现裴氏着色霉具有一定的种内差异 ,其DNA带型与菌株的来源地域关系密切 ,本研究方法简便快速 ,可用于着色芽生菌病的分子流行病学调查及可能的基因学诊断中。
Objective To further understand the genetic characteristics of Peyer’s stain mildew, extract genomic DNA with bromobenzene extraction. Methods Random amplified polymorphic DNA (PCR) was used to detect 32 species of Peyer’s patches (7 Japanese, 3 Chinese, 4 Venezuelan, 4 Central American, 2 Colombian, 2 Mexican, Brazil 1 strain, Thailand 1 strain) DNA fingerprinting was analyzed. Results ① A total of 10 random primers were selected for amplification. Three primers with better DNA amplification were screened: primer 1: 5 ’GAGCC CTCCA 3’, primer 7: 5 ’CGGCCCCTGT 3’, primer 8: 5 ’AGGTCACTGA 3’. (2) The DNA patterns of 3 2 Peyer’s patches were not exactly the same, with some genetic variability. ③ strains from the same country have a higher genetic similarity. (4) There were two types of DNA bands in the 15 strains derived from Japan, including type I (three major bands) and type II (two major bands), and the rest of the national strains were small in number. DNA Sub-type is not obvious. ⑤ amplification of the same primer, more than 90% of strains can be common DNA fragments. Conclusion The results of random amplified polymorphic DNA (DNA) polymorphism study showed that there are some intraspecific differences in P. chrysogenum, and their DNA bands are closely related to the origin of the strains. The method is simple and rapid and can be applied to the molecular epidemiology of colored blastomycosis Investigate and possible genetic diagnosis.