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逆转座子-微卫星扩增多态性,一种研究日本血吸虫地理株遗传变异的电泳方法。

Retrotransposon-microsatellite amplified polymorphism, an electrophoretic approach for studying genetic variability among Schistosoma japonicum geographical isolates.

机构信息

Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province, People's Republic of China.

出版信息

Electrophoresis. 2012 Sep;33(18):2859-66. doi: 10.1002/elps.201200110.

Abstract

In the present study, retrotransposon-microsatellite amplified polymorphism (REMAP) was used to examine genetic variability among Schistosoma japonicum isolates from different endemic provinces in mainland China, using S. japonicum from Japan and the Philippines for comparison. Of the 50 primer combinations screened, eight produced highly reproducible REMAP fragments. Using these primers, 190 distinct DNA fragments were generated in total, of which 147 (77.37%) were polymorphic, indicating considerable genetic variation among the 43 S. japonicum isolates examined. The percentage of polymorphic bands (PPB) among S. japonicum isolates from mainland China, Japan, and the Philippines was 77.37%; PPB values of 18.42% and 53.68% were found among isolates from southwestern (SW) China and the lower Yangtze/Zhejiang province in eastern (E) China, respectively. Based on REMAP profiles, unweighted pair-group method with arithmetic averages (UPGMA) dendrogram analysis revealed that all of the S. japonicum samples grouped into three distinct clusters: parasites from mainland China, Japan, and the Philippines were clustered in each individual clade. Within the mainland China cluster, SW China isolates (from Sichuan and Yunnan provinces) grouped together, whereas worms from E China (Zhejiang, Anhui, Jiangxi, Jiangsu, Hunan, and Hubei provinces) grouped together. These results demonstrated that the REMAP marker system provides a reliable electrophoretic technique for studying genetic diversity and population structures of S. japonicum isolates from mainland China, and could be applied to other pathogens of human and animal health significance.

摘要

在本研究中,我们使用逆转座子-微卫星扩增多态性(REMAP)技术,以日本和菲律宾的血吸虫作为对照,检测中国大陆不同流行省份的日本血吸虫分离株的遗传变异。在筛选的 50 对引物组合中,有 8 对产生了高度可重复的 REMAP 片段。使用这些引物,总共产生了 190 个不同的 DNA 片段,其中 147 个(77.37%)为多态性,表明所检测的 43 个日本血吸虫分离株存在相当大的遗传变异。中国大陆、日本和菲律宾的日本血吸虫分离株的多态带百分比(PPB)分别为 77.37%;中国西南部(SW)和华东(E)长江中下游/浙江的分离株的 PPB 值分别为 18.42%和 53.68%。基于 REMAP 图谱,非加权对组平均法(UPGMA)聚类分析显示,所有日本血吸虫样本分为三个明显的聚类:中国大陆、日本和菲律宾的寄生虫分别聚类在各自的分支中。在中国大陆的聚类中,SW 中国的分离株(来自四川和云南省)聚在一起,而来自 E 中国(浙江、安徽、江西、江苏、湖南和湖北省)的蠕虫聚在一起。这些结果表明,REMAP 标记系统为研究中国大陆日本血吸虫分离株的遗传多样性和种群结构提供了一种可靠的电泳技术,并且可以应用于其他人类和动物健康重要病原体。

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