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Molecular Identification, Transcriptome Sequencing and Functional Annotation of Pulex irritans
Acta Parasitologica ( IF 1.5 ) Pub Date : 2021-01-03 , DOI: 10.1007/s11686-020-00296-x
Li Hu 1 , Yae Zhao 1 , Yanan Yang 1 , Wanyu Zhang 1 , Hongsong Guo 1 , Dongling Niu 1
Affiliation  

Purpose

Pulex irritans are vectors of various zoonotic pathogens. However, molecular studies on P. irritans and flea-borne diseases are limited due to the lack of molecular data. This study aimed to conduct transcriptome sequencing, functional annotation, and pathogen analysis of P. irritans.

Methods

Fleas collected from a dog were identified morphologically and molecularly. RNA was extracted for transcriptome sequencing and functional annotation. Open reading frames (ORFs) of unigenes were confirmed by employing bioinformatics strategies, and maximum likelihood (ML) trees were reconstructed based on the highly expressed genes of ejaculation globulin-specific 3-like protein, salivary protein, and actin for phylogenetic relationship analysis.

Results

The obtained mitochondrial 16S rRNA gene sequences showed 99.71% of similarity with P. irritans obtained from GenBank database. Transcriptome sequencing generated 74,412 unigenes, of which 53,211 were functionally annotated. A total of 195 unigenes were assigned to fleas, of which 69 contained complete ORFs. Phylogenetic trees of both ejaculatory globulin and salivary protein genes demonstrated that P. irritans first clustered with Pulicidae sp., indicating the reliability of transcriptome data. It is noteworthy that 1070 unigenes were assigned to Hymenolepis microstoma and Dipylidium caninum, of which 62 contained complete ORFs. The phylogenetic tree of the actin gene showed that the unigenes had closer relationships with Echinococcus sp., suggesting the role of P. irritans as intermediate hosts of tapeworms.

Conclusion

The results of this study provide the possibility for functional exploration of important genes and lay foundations for the prevention and control of P. irritans and flea-borne diseases.



中文翻译:

Pulex irritans 的分子鉴定、转录组测序和功能注释

目的

Pulex irritans是各种人畜共患病原体的载体。然而,由于缺乏分子数据,对刺激性疟原虫和跳蚤传播疾病的分子研究受到限制。本研究旨在对刺激性疟原虫进行转录组测序、功能注释和病原体分析。

方法

从狗身上收集的跳蚤在形态学和分子学上都得到了鉴定。提取 RNA 用于转录组测序和功能注释。采用生物信息学策略确认unigenes的开放阅读框(ORFs),并基于射精球蛋白特异性3样蛋白、唾液蛋白和肌动蛋白的高表达基因重建最大似然(ML)树进行系统发育关系分析。

结果

获得的线粒体16S rRNA基因序列与GenBank数据库中的P. irritans具有99.71%的相似性。转录组测序生成了 74,412 个 unigenes,其中 53,211 个进行了功能注释。共有 195 个 unigenes 被分配给跳蚤,其中 69 个包含完整的 ORF。射精球蛋白和唾液蛋白基因的系统发育树表明,P. irritans首先与Pulicidae sp.聚类,表明转录组数据的可靠性。值得注意的是,1070个unigenes被分配到Hymenolepis microstomaDipylidium caninum,其中 62 个包含完整的 ORF。肌动蛋白基因的系统发育树表明,unigenes 与棘球绦虫的关系更密切,表明P. irritans作为绦虫的中间宿主。

结论

本研究结果为重要基因的功能探索提供了可能,为防治刺激性疟原虫和跳蚤传播疾病奠定了基础。

更新日期:2021-01-03
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