硬蜱属基因在吸血和长期饥饿后差异表达的转录组从头组装和分析。
De novo assembly and analysis of the transcriptome of the Dermacentor marginatus genes differentially expressed after blood-feeding and long-term starvation.
机构信息
College of Animal Science, Xinjiang Agricultural University, Ürümqi, 830052, Xinjiang Uygur Autonomous Region, People's Republic of China.
College of Veterinary Medicine, Xinjiang Agricultural University, Ürümqi, 830052, Xinjiang Uygur Autonomous Region, People's Republic of China.
出版信息
Parasit Vectors. 2020 Nov 10;13(1):563. doi: 10.1186/s13071-020-04442-2.
BACKGROUND
The ixodid tick Dermacentor marginatus is a vector of many pathogens wide spread in Eurasia. Studies of gene sequence on many tick species have greatly increased the information on tick protective antigen which might have the potential to function as effective vaccine candidates or drug targets for eco-friendly acaricide development. In the current study, RNA-seq was applied to identify D. marginatus sequences and analyze differentially expressed unigenes.
METHODS
To obtain a broader picture of gene sequences and changes in expression level, RNA-seq was performed to obtain the whole-body transcriptome data of D. marginatus adult female ticks after engorgement and long-term starvation. Subsequently, the real-time quantitative PCR (RT-qPCR) was applied to validate the RNA-seq data.
RESULTS
RNA-seq produced 30,251 unigenes, of which 32% were annotated. Gene expression was compared among groups that differed by status as newly molted, starved and engorged female adult ticks. Nearly one third of the unigenes in each group were differentially expressed compared to the other two groups, and the most numerous were genes encoding proteins involved in catalytic and binding activities and apoptosis. Selected up-regulated differentially expressed genes in each group were associated to protein, lipids, carbohydrate and chitin metabolism. Blood-feeding and long-term starvation also caused genes differentially expressed in the defense response and antioxidant response. RT-qPCR results indicated 6 differentially expressed transcripts showed similar trends in expression changes with RNA-seq results confirming that the gene expression profiles in transcriptome data is in consistent with RT-qPCR validation.
CONCLUSIONS
Obtaining the sequence information of D. marginatus and characterizing the expression pattern of the genes involved in blood-feeding and during starvation would be helpful in understanding molecular physiology of D. marginatus and provides data for anti-tick vaccine and drug development for controlling the tick.
背景
嗜群血蜱 Dermacentor marginatus 是一种广泛分布于欧亚大陆的许多病原体的媒介。对许多蜱物种的基因序列研究极大地增加了蜱保护性抗原的信息，这些信息可能具有作为有效疫苗候选物或药物靶点的潜力，以用于生态友好型杀蜱剂的开发。在本研究中，我们应用 RNA-seq 来鉴定 Dermacentor marginatus 的序列并分析差异表达的基因。
方法
为了更全面地了解基因序列和表达水平的变化，我们进行了 RNA-seq 实验，以获得饱血和长期饥饿后 Dermacentor marginatus 成年雌蜱的全虫转录组数据。随后，我们应用实时定量 PCR（RT-qPCR）来验证 RNA-seq 数据。
结果
RNA-seq 产生了 30251 条 unigenes，其中 32%被注释。我们比较了新蜕皮、饥饿和饱血的雌蜱之间的基因表达。与其他两组相比，每组中有近三分之一的 unigenes差异表达，其中数量最多的是参与催化和结合活性以及细胞凋亡的蛋白质编码基因。每个组中上调的差异表达基因与蛋白质、脂质、碳水化合物和几丁质代谢有关。吸血和长期饥饿也导致了防御反应和抗氧化反应中的基因差异表达。RT-qPCR 结果表明，6 个差异表达的转录本在表达变化上表现出与 RNA-seq 结果相似的趋势，这证实了转录组数据中的基因表达谱与 RT-qPCR 验证结果一致。
结论
获得 Dermacentor marginatus 的序列信息并描述参与吸血和饥饿过程中的基因表达模式有助于理解 Dermacentor marginatus 的分子生理学，并为抗蜱疫苗和药物开发提供数据，以控制蜱的数量。
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