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[中国大陆土耳其斯坦东毕吸虫rDNA-LSU基因的序列分析]

[Sequence analysis of rDNA-LSU gene of Orientobilharzia turkestanicum from mainland of China].

作者信息

Zhang Guang-jun, Chen Qin, Qiu Chi-ping, Xia Ming-yi

机构信息

Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai 200025.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2002;20(2):86-9.

Abstract

OBJECTIVE

To classify the taxonomic status of O. cheni in relation to O. turkestanicum var. tuberculata from the mainland of China by comparing their nucleotide sequences of nuclear ribosomal partial large subunit gene (LSU).

METHODS

The genomic DNA of adult worms were extracted by the GNT-K method. The target gene was amplified by PCR using specific primers. The PCR products were purified before ligation into the plasmid PCR-blunt (Invitrogen). Recombinant plasmids were amplified in E. coli, extracted and purified using routine methods and then sequenced using M13 primers (F/R) on a Licor long-read auto-sequencer. Sequences of O. turkestanicum was retrieved from GenBank and aligned with our data in BioEdit.

RESULTS

The nucleotide sequences of LSU between O. turkestanicum var. tuberculata and O. cheni was 100% identical, and 99.99% identical between O. turkestanicum var. tuberculata and O. turkestanicum.

CONCLUSION

This study demonstrated high similarity in LSU nucleotide sequences, and the results do not support O. cheni as an independent species. O. cheni may be a synonym of O. turkestanicum var. tuberculata, and O. turkestanicum var. tuberculata is probably also a synonym of O. turkestanicum.

摘要

目的

通过比较陈氏奥斯特线虫与中国内地粗纹奥斯特线虫变种的核糖体大亚基基因(LSU)部分核苷酸序列,对陈氏奥斯特线虫的分类地位进行界定。

方法

采用GNT-K法提取成虫基因组DNA。使用特异性引物通过PCR扩增目标基因。PCR产物在连接到质粒PCR-blunt(Invitrogen)之前进行纯化。重组质粒在大肠杆菌中扩增,采用常规方法提取和纯化,然后使用M13引物(F/R)在Licor长读长自动测序仪上进行测序。从GenBank中检索粗纹奥斯特线虫的序列,并在BioEdit中与我们的数据进行比对。

结果

粗纹奥斯特线虫变种与陈氏奥斯特线虫之间LSU的核苷酸序列完全相同,粗纹奥斯特线虫变种与粗纹奥斯特线虫之间的序列相似度为99.99%。

结论

本研究表明LSU核苷酸序列具有高度相似性,结果不支持陈氏奥斯特线虫为独立物种。陈氏奥斯特线虫可能是粗纹奥斯特线虫变种的同义词,而粗纹奥斯特线虫变种可能也是粗纹奥斯特线虫的同义词。

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