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条纹鲈(Morone saxatilis)是一种高度进化的鲈形目鱼类,该研究构建了其所有成熟阶段的卵巢转录组。

An ovary transcriptome for all maturational stages of the striped bass (Morone saxatilis), a highly advanced perciform fish.

作者信息

Reading Benjamin J, Chapman Robert W, Schaff Jennifer E, Scholl Elizabeth H, Opperman Charles H, Sullivan Craig V

机构信息

North Carolina State University, Department of Biology, Raleigh, NC, USA.

出版信息

BMC Res Notes. 2012 Feb 21;5:111. doi: 10.1186/1756-0500-5-111.

Abstract

BACKGROUND

The striped bass and its relatives (genus Morone) are important fisheries and aquaculture species native to estuaries and rivers of the Atlantic coast and Gulf of Mexico in North America. To open avenues of gene expression research on reproduction and breeding of striped bass, we generated a collection of expressed sequence tags (ESTs) from a complementary DNA (cDNA) library representative of their ovarian transcriptome.

RESULTS

Sequences of a total of 230,151 ESTs (51,259,448 bp) were acquired by Roche 454 pyrosequencing of cDNA pooled from ovarian tissues obtained at all stages of oocyte growth, at ovulation (eggs), and during preovulatory atresia. Quality filtering of ESTs allowed assembly of 11,208 high-quality contigs ≥ 100 bp, including 2,984 contigs 500 bp or longer (average length 895 bp). Blastx comparisons revealed 5,482 gene orthologues (E-value < 10-3), of which 4,120 (36.7% of total contigs) were annotated with Gene Ontology terms (E-value < 10-6). There were 5,726 remaining unknown unique sequences (51.1% of total contigs). All of the high-quality EST sequences are available in the National Center for Biotechnology Information (NCBI) Short Read Archive (GenBank: SRX007394). Informative contigs were considered to be abundant if they were assembled from groups of ESTs comprising ≥ 0.15% of the total short read sequences (≥ 345 reads/contig). Approximately 52.5% of these abundant contigs were predicted to have predominant ovary expression through digital differential display in silico comparisons to zebrafish (Danio rerio) UniGene orthologues. Over 1,300 Gene Ontology terms from Biological Process classes of Reproduction, Reproductive process, and Developmental process were assigned to this collection of annotated contigs.

CONCLUSIONS

This first large reference sequence database available for the ecologically and economically important temperate basses (genus Morone) provides a foundation for gene expression studies in these species. The predicted predominance of ovary gene expression and assignment of directly relevant Gene Ontology classes suggests a powerful utility of this dataset for analysis of ovarian gene expression related to fundamental questions of oogenesis. Additionally, a high definition Agilent 60-mer oligo ovary 'UniClone' microarray with 8 × 15,000 probe format has been designed based on this striped bass transcriptome (eArray Group: Striper Group, Design ID: 029004).

摘要

背景

条纹鲈及其近缘种(Morone属)是重要的渔业和水产养殖物种,原产于北美大西洋沿岸和墨西哥湾的河口及河流。为了开辟条纹鲈繁殖和育种方面的基因表达研究途径,我们从一个代表其卵巢转录组的互补DNA(cDNA)文库中生成了一组表达序列标签(EST)。

结果

通过对从卵母细胞生长各阶段、排卵(卵子)时以及排卵前闭锁期获得的卵巢组织混合cDNA进行罗氏454焦磷酸测序,共获得了230,151个EST序列(51,259,448碱基对)。对EST进行质量过滤后,组装得到了11,208个长度≥100碱基对的高质量重叠群,其中包括2,984个长度为500碱基对或更长的重叠群(平均长度895碱基对)。Blastx比对揭示了5,482个基因直系同源物(E值<10^-3),其中4,120个(占重叠群总数的36.7%)被赋予了基因本体论术语(E值<10^-6)。还剩下5,726个未知的独特序列(占重叠群总数的51.1%)。所有高质量的EST序列都可在国家生物技术信息中心(NCBI)的短读存档库中获取(GenBank:SRX007394)。如果信息性重叠群是由占总短读序列≥0.15%(≥345条读段/重叠群)的EST组组装而成,则被认为是丰富的。通过与斑马鱼(Danio rerio)单基因直系同源物进行电子差异显示比较,预计这些丰富重叠群中约52.5%在卵巢中具有优势表达。超过1300个来自生殖、生殖过程和发育过程等生物学过程类别的基因本体论术语被赋予了这组注释重叠群。

结论

这个首个可用于生态和经济上重要的温带鲈鱼(Morone属)的大型参考序列数据库为这些物种的基因表达研究奠定了基础。预测的卵巢基因表达优势以及直接相关的基因本体论类别的赋予表明,该数据集对于分析与卵子发生基本问题相关的卵巢基因表达具有强大的实用性。此外,基于这个条纹鲈转录组设计了一种具有8×15,000探针格式的高分辨率安捷伦60聚体卵巢“单克隆”微阵列(eArray组:条纹鲈组,设计ID:029004)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/41d8/3305648/5e6601f8f4ee/1756-0500-5-111-1.jpg

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