PDF(Pubmed)
Abstract：
To conduct differential gene expression analysis, ovaries from the cattle tick Rhipicephalus microplus were dissected at three distinct developmental stages (preingurgitated, immature ingurgitated, and mature ingurgitated). Additionally, undissected intact mature males and complete ingurgitated female ticks without ovaries (carcasses) were also collected to serve as reference samples for analysis. To perform total RNA purification, tissue from ten individuals representing each of the five previously described conditions was pooled. mRNA was isolated from the purified total RNA using the oligo (dT) method. Following fragmentation, double stranded cDNA was synthesized and ligated to sequencing adapters. Suitable-sized fragments were subsequently used for PCR amplification. Libraries were analyzed and quantified using an Agilent 2100 Bioanalyzer and an ABI StepOnePlus Real-Time PCR System. A total of 45.64 Gb bases were sequenced using the Illumina HiSeq sequencing platform. After assembling the samples and correcting for abundance, we obtained 82,877 unigenes. The total length, average length, N50, and GC content of the unigenes were 89,754,828 bp,1,082 bp,2,068 bp and 49.04 % respectively. For functional annotation, the unigenes were aligned with 7 functional databases. The number of unigenes identified in the functional databases were as follows: 32,518 (NR:39.24 %), 10,259 (NT:12.38 %), 23,624 (Swissprot:28.50 %), 22,203 (KOG:26.79 %), 25,072 (KEGG:30.25 %), 17,435(GO:21.04 %), and 23,220 (InterPro:28.02 %). Unigene candidate coding regions (CDS) among the unigenes were predicted using TransDecoder software and 42,143 CDS were detected. We also detected 10,522 simple sequence repeats (SSRs) distributed on 8,126 unigenes, and predicted 4,672 transcription factors (TF) coding unigenes. Our data can be used to identify genes that are important for male and female tick and arachnid reproduction and tick general physiology.
摘要：
进行差异基因表达分析,在三个不同的发育阶段解剖了来自牛tickRhipicephalusmicroplus的卵巢（前，不成熟的，和成熟的人)。此外,还收集了未解剖的完整成熟雄性和没有卵巢（car体）的完整吞噬雌性蜱，作为分析的参考样品。要进行总RNA纯化，汇集了来自10个个体的组织,这些个体代表了前述5个条件中的每一个。使用oligo(dT)方法从纯化的总RNA中分离mRNA。在碎片化之后，合成双链cDNA并连接到测序衔接子。随后将合适大小的片段用于PCR扩增。使用Agilent2100生物分析仪和ABIStepOnePlus实时PCR系统分析和定量文库。使用IlluminaHiSeq测序平台对总共45.64Gb碱基进行测序。组装样品并校正丰度后，我们获得了82,877个基因。总长度,平均长度,N50和GC含量为89,754,828bp，1,082bp，分别为2,068bp和49.04%。对于功能注释，单基因与7个功能数据库进行比对.功能数据库中鉴定的单基因数量如下：32,518（NR：39.24％），10,259（新台币：12.38%），23,624（Swissprot：28.50%），22,203（KOG：26.79％），25,072（KEGG：30.25％），17,435（GO：21.04%），和23,220（InterPro：28.02%）。使用TransDecoder软件预测单基因中的单基因候选编码区（CDS），并检测到42,143个CDS。我们还检测到分布在8,126个单基因上的10,522个简单序列重复(SSR)，并预测了4,672个转录因子（TF）编码单基因。我们的数据可用于鉴定对雄性和雌性蜱和蜘蛛繁殖以及蜱一般生理学重要的基因。
