Sequencing and analysis on transcriptomes of unfed female Haemaphysalis longicornis

Abstract

Objective To sequence transcriptomes of unfed female Haemaphysalis longicornis using IlluminaHiSeq highthroughput technology. Methods The data on sequences the transcriptomes were spliced and assembled, and the obtained sequences were analyzed with functional annotation, functional classification, metabolic pathway analysis and simple repeated sequence markers using bioinformatics methods. Results A total of 181 246 184 clean reads data were obtained and 107 428 unigene sequences were obtained after assembly, with an average length of 1 246.29. All unigene sequences were aligned with the Nr, Nt, Pfam, KOG, Swiss-prot, KEGG, GO databases using BLAST software. Compared with the Nr database, the long-horned blood scorpion gene sequence has a high homology (55.3%) with that of Ixodes scapularis . According to the annotation of the GO database, the functions of the all unigene sequences were divided into 3 categories (biological process, cellular component and molecular function) covering 56 functional groups; based on to the annotations of the KOG database, all the unigene sequences were assigned into 25 categories; while, according to the analysis of the KEGG database, there are 32 groups of genes involved in metabolic pathways and a major part of them (12.13%) are involved in signal transduction. A total of 45 863 simple sequence repeats (SSRs) were identified with SSR locus search. Single nucleotide polymorphism (SNP) analysis indicated that the number of SNPs for base transition was 195 369 and that for base transversion was 96 780. Conclusion The analysis on transcriptomes of unfed female Haemaphysalis longicornis lays a foundation for subsequent researches on gene expression and expression of the tick 【摘 要】 目的 利用 IlluminaHiSeq 高通量技术对长角血蜱饥饿雌成蜱进行转录组测序。 方法 将测序得到的数据进行拼接组装，并利用生物信息学方法对所得序列进行基因功能注释、功能分类、代谢途径分析和简单重复序列标记等分析。 结果 共获得 181 246 184 个 clean reads 数据，组装后得到 107 428 个 unigenes 序列，平均长度 1 246.29。以 BLAST 软件将所有的 unigenes 序列与 Nr，Nt，Pfam，KOG，Swiss-prot，KEGG，GO 数据库进行比对。与 Nr 数据库比对发现长角血蜱基因序列与同科肩突硬蜱（ Ixodes scapularis ）具有较高的同源性，为 55.3 %；根据 GO 数据库的注释，将其分为生物过程、细胞组分和分子功能 3 大类共 56 分支；通过与 KOG 数据库的注释划分 25 类；根据与 KEGG 数据库的分析发现含有代谢通路相关的基因有 32 类，其中较多的是信号转导（signal transduction）类，占 12.13 %。通过 SSR 位点查找共鉴定出 45 863 个 SSR；SNP 分析发现发生转换的 SNP 数为 195 369 个，发生颠换的 SNP 数为 96 780 个。 结论 通过对长角血蜱饥饿雌成蜱转录组水平分析，可为后续长角血蜱功能基因的挖掘及表达谱等方面的研究奠定基础。