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构建并鉴定鲨鱼再生肝组织的 cDNA 文库。

Construction and characterization of a cDNA library from shark regenerated hepatic tissue.

机构信息

School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, PR China.

出版信息

Fish Shellfish Immunol. 2011 Apr-May;30(4-5):1170-7. doi: 10.1016/j.fsi.2011.02.025. Epub 2011 Mar 5.

Abstract

Sharks are a type of fish with a full cartilaginous skeleton and have big livers. To better understand liver regeneration in sharks and screening for the important genes participated in disease-defense, in this study, a cDNA library of regenerated liver tissues of shark, Chiloscyllium plagiosum, was constructed. A total of 2103 expressed sequence tags (ESTs), which represents 997 unique genes, were sequenced. Among these genes, 434 (43.53%) of them showed significant similarity (E-values < 10⁻⁵) to the sequences in NCBI Nt database, 685 (68.71%) of these unique genes showed significant similarity (E-values < 10⁻⁵) to the sequences in NCBI Nr database, and 662 (66.40%) of these unique genes showed significant similarity (E-values < 10⁻⁵) to the Swiss-Prot database. Preliminary analysis of unique genes according to COG database showed that unigenes were further grouped into 21 functional categories including inorganic ion transport and metabolism, energy production and conversion, posttranslational modification, protein turnover and chaperones, general function prediction only, translation, and ribosomal structure and biogenesis. Several possible candidate genes involved in liver regeneration were selected to analyze their expression with relative quantification real-time PCR. This study may contribute to our better understanding of the molecular mechanism of regeneration in shark liver. Furthermore, the EST cataloguing and profiling of shark will be also benefited to the functional genomic research in this marine species.

摘要

鲨鱼是一种具有完全软骨骨架的鱼类,其肝脏较大。为了更好地了解鲨鱼肝脏的再生以及筛选参与疾病防御的重要基因,本研究构建了软骨鲨(Chiloscyllium plagiosum)再生肝脏组织的 cDNA 文库。共测序了 2103 个表达序列标签(EST),代表 997 个独特基因。在这些基因中,有 434 个(43.53%)与 NCBI Nt 数据库中的序列具有显著相似性(E 值 < 10⁻⁵),685 个(68.71%)与 NCBI Nr 数据库中的序列具有显著相似性(E 值 < 10⁻⁵),662 个(66.40%)与 Swiss-Prot 数据库中的序列具有显著相似性(E 值 < 10⁻⁵)。根据 COG 数据库对独特基因进行初步分析表明,unigenes 进一步分为 21 个功能类别,包括无机离子运输和代谢、能量产生和转化、翻译后修饰、蛋白质周转和伴侣、一般功能预测仅、翻译和核糖体结构和生物发生。选择了几个可能参与肝脏再生的候选基因,通过相对定量实时 PCR 分析它们的表达。本研究有助于我们更好地了解鲨鱼肝脏再生的分子机制。此外,对鲨鱼的 EST 编目和分析也将有助于该海洋物种的功能基因组研究。

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