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柔嫩艾美耳球虫不同发育阶段的比较转录组分析及在孢子生殖阶段特异性表达的一个 ApiAP2 转录因子的功能分析。

Comparative transcriptome profiling of Eimeria tenella in various developmental stages and functional analysis of an ApiAP2 transcription factor exclusively expressed during sporogony.

机构信息

National Key Laboratory of Veterinary Public Health Security, Key Laboratory of Animal Epidemiology and Zoonosis of Ministry of Agriculture, National Animal Protozoa Laboratory & College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China.

Key Laboratory of Animal Biosafety Risk Prevention and Control (North) of MARA, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

出版信息

Parasit Vectors. 2023 Jul 19;16(1):241. doi: 10.1186/s13071-023-05828-8.

Abstract

BACKGROUND

The apicomplexan parasites Eimeria spp. are the causative agents of coccidiosis, a disease with a significant global impact on the poultry industry. The complex life cycle of Eimeria spp. involves exogenous (sporogony) and endogenous (schizogony and gametogony) stages. Unfortunately, the genetic regulation of these highly dynamic processes, particularly for genes involved in specific developmental phases, is not well understood.

METHODS

In this study, we used RNA sequencing (RNA-Seq) analysis to identify expressed genes and differentially expressed genes (DEGs) at seven time points representing different developmental stages of Eimeria tenella. We then performed K-means clustering along with co-expression analysis to identify functionally enriched gene clusters. Additionally, we predicted apicomplexan AP2 transcription factors in E. tenella using bioinformatics methods. Finally, we generated overexpression and knockout strains of ETH2_0411800 to observe its impact on E. tenella development.

RESULTS

In total, we identified 7329 genes that are expressed during various developmental stages, with 3342 genes exhibiting differential expression during development. Using K-means clustering along with co-expression analysis, we identified clusters functionally enriched for oocyte meiosis, cell cycle, and signaling pathway. Among the 53 predicted ApiAP2 transcription factors, ETH2_0411800 was found to be exclusively expressed during sporogony. The ETH2_0411800 overexpression and knockout strains did not exhibit significant differences in oocyst size or output compared to the parental strain, while the resulting ETH2_0411800 knockout parasite showed a relatively small oocyst output.

CONCLUSIONS

The findings of our research suggest that ETH2_0411800 is not essential for the growth and development of E. tenella. Our study provides insights into the gene expression dynamics and is a valuable resource for exploring the roles of transcription factor genes in regulating the development of Eimeria parasites.

摘要

背景

艾美耳球虫属的顶复门寄生虫是球虫病的病原体,这种疾病对全球家禽养殖业有重大影响。艾美耳球虫属的复杂生命周期包括外生性(孢子生殖)和内生性(裂殖生殖和配子生殖)阶段。不幸的是,这些高度动态过程的遗传调控,特别是涉及特定发育阶段的基因,还没有得到很好的理解。

方法

在这项研究中,我们使用 RNA 测序(RNA-Seq)分析来鉴定七个时间点代表艾美耳球虫发育不同阶段的表达基因和差异表达基因(DEGs)。然后,我们进行了 K-means 聚类和共表达分析,以鉴定功能丰富的基因簇。此外,我们使用生物信息学方法预测了艾美耳球虫中的顶复门 AP2 转录因子。最后,我们生成了 ETH2_0411800 的过表达和敲除株,以观察其对艾美耳球虫发育的影响。

结果

我们总共鉴定了 7329 个在不同发育阶段表达的基因,其中 3342 个基因在发育过程中表现出差异表达。通过 K-means 聚类和共表达分析,我们鉴定了功能丰富的卵母细胞减数分裂、细胞周期和信号通路聚类。在预测的 53 个 ApiAP2 转录因子中,ETH2_0411800 仅在孢子生殖阶段表达。与亲本株相比,ETH2_0411800 的过表达和敲除株在卵囊大小或产量方面没有显著差异,而产生的 ETH2_0411800 敲除寄生虫的卵囊产量相对较小。

结论

我们的研究结果表明,ETH2_0411800 对于艾美耳球虫的生长和发育不是必需的。我们的研究提供了对基因表达动态的深入了解,是探索转录因子基因在调节艾美耳球虫发育中的作用的宝贵资源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d404/10354945/f5548470564f/13071_2023_5828_Fig1_HTML.jpg

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