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家鸡艾美耳球虫:利用任意引物扩增的DNA多态性和变性梯度凝胶电泳分析种间及株间的遗传变异性

Eimeria spp. of the domestic fowl: analysis of genetic variability between species and strains using DNA polymorphisms amplified by arbitrary primers and denaturing gradient-gel electrophoresis.

作者信息

Johnston D A, Fernando M A

机构信息

Department of Pathology, University of Guelph, Ontario, Canada.

出版信息

Parasitol Res. 1995;81(2):91-7. doi: 10.1007/BF00931611.

Abstract

The genetic relatedness of 5 Eimeria spp. of the domestic fowl, including 11 strains of E. acervulina, 2 strains of E. tenella and 1 precocious line of E. acervulina, was assayed by means of random amplified polymorphic DNA (RAPD) and denaturing gradient-gel electrophoresis (DGGE). Seven different oligonucleotides were used to generate similarity coefficients for the species and strains of Eimeria infecting chickens. Between 1 and 13 DNA segments, depending on the species/strain-primer combination, were amplified with the various primers. Amplification products ranged in size from 0.16 to 3.8 kb. E. acervulina strains demonstrated two to four major common bands unique to the species. These strains also exhibited major and minor differences in their DNA patterns. Band-match analyses from both polyacrylamide and denaturing gradient gels were used to calculate similarity coefficients for the Eimeria spp. and strains tested. Species differences, readily detected upon examination of DNA banding patterns, gave similarity coefficients of 4%-38% and 3%-18% when analyzed by polyacrylamide- and denaturing gradient-gel electrophoresis, respectively. A similar analysis of E. acervulina strains, yielded similarity coefficients of 55%-95% and 51%-85%, respectively. The differences observed between both species and strains were greater when the RAPD-assay products were analyzed via DGGE, indicating that a combination of these two techniques may provide a more stringent analysis of the genetic relatedness of these coccidia.

摘要

利用随机扩增多态性DNA(RAPD)和变性梯度凝胶电泳(DGGE)技术,对家鸡的5种艾美耳球虫的遗传相关性进行了分析,其中包括11株堆型艾美耳球虫、2株柔嫩艾美耳球虫和1个堆型艾美耳球虫早熟系。使用7种不同的寡核苷酸来生成感染鸡的艾美耳球虫物种和菌株的相似系数。根据物种/菌株 - 引物组合的不同,各种引物扩增出1至13个DNA片段。扩增产物的大小在0.16至3.8 kb之间。堆型艾美耳球虫菌株显示出该物种特有的2至4条主要共有带。这些菌株在DNA图谱上也表现出主要和次要的差异。利用聚丙烯酰胺凝胶和变性梯度凝胶的条带匹配分析来计算所测试的艾美耳球虫物种和菌株的相似系数。通过检查DNA条带模式很容易检测到物种差异,当通过聚丙烯酰胺凝胶电泳和变性梯度凝胶电泳分析时,相似系数分别为4% - 38%和3% - 18%。对堆型艾美耳球虫菌株进行的类似分析,相似系数分别为55% - 95%和51% - 85%。当通过DGGE分析RAPD测定产物时,观察到的物种和菌株之间的差异更大,这表明这两种技术的结合可能会对这些球虫的遗传相关性提供更严格的分析。

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