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应用核糖体DNA(ribo-somal DNAr,DNA)的ITS(internal transcribed spacer,ITS)区的RFLP序列的多态性,分析32个不同寄主来源的炭疽菌株的遗传多样性。利用ITS1及ITS4通用引物对扩增各菌株的rD-NA的内转录间隔区(ITS1-5.8S-ITS2)获得长约600 bp片段。ITS-RFLP共迁带UPGMA聚类分析的结果表明所有来自不同寄主的炭疽菌菌株(含福建及云南菌株)都以较高的相似系数(>0.60)聚为一类,具有较近的亲缘关系;同时试验中所有酶切位点的综合Gst系数达到0.82,说明不同菌株间的ITS具有丰富的多态性。
The genetic diversity of anthrax strains from 32 different host plants was analyzed by using the RFLP sequence polymorphism of ITS (internal transcribed spacer ITS) region of ribosome DNA (DNA). The ITS1-5.8S-ITS2 was amplified by using ITS1 and ITS4 universal primers to amplify the rD-NA internal transcribed spacer of each strain, and a 600 bp fragment was obtained. The results of UPGMA clustering analysis showed that all the anthrax strains (including Fujian and Yunnan strains) from different hosts clustered together with higher similarity coefficients (> 0.60) and had closer genetic relationship At the same time, the overall Gst coefficient of all the enzyme cutting sites in the experiment reached 0.82, indicating that ITS among different strains is rich in polymorphism.