文献类型：期刊文献

中文题名：溶藻弧菌诱导马氏珠母贝血淋巴cDNA差减文库的构建及分析

英文题名：Analysis of differentially expressed genes in response to Vibrio alginolyticus stimulation in hemocytes of pearl oyster( Pinctada fucata)

作者：王忠良[1,2];简纪常[1,3];鲁义善[1,3];王蓓[1,3];陈刚[1,2];吴灶和[3,4]

机构：[1]广东海洋大学水产学院,广东湛江524088;[2]广东海洋大学南海水产经济动物增养殖重点实验室,广东湛江524088;[3]广东省水产经济动物病原生物学及流行病学重点实验室,广东湛江524088;[4]仲恺农业工程学院,广东广州510225

年份：2014

卷号：38

期号：1

起止页码：127

中文期刊名：水产学报

外文期刊名：Journal of Fisheries of China

收录：CSTPCD、、北大核心2011、Scopus、北大核心、CSCD、CSCD2013_2014

基金：国家自然科学基金项目(31202023)

语种：中文

中文关键词：马氏珠母贝;溶藻弧菌;抑制性消减杂交;cDNA文库

外文关键词：Pinctada fucata ; Vibrio alginolyticus ; SSH ; cDNA library

中文摘要：为了探讨马氏珠母贝免疫防御机制，筛选免疫防御相关功能基因，以致病性溶藻弧菌人工感染的马氏珠母贝血淋巴为材料，采用抑制性差减杂交（SSH）技术，构建了溶藻弧菌诱导的马氏珠母贝血淋巴cDNA差减文库；以马氏珠母贝管家基因β-actin作为差减指标检测该文库的差减效率；对选取的600个阳性克隆进行测序及生物信息学分析。结果显示，该文库的差减效率可达2^10倍；PCR阳性检测显示差减片段为100～750bp，对随机挑取600个克隆的测序结果显示，共获得414个有效EST序列；通过BLAST同源性比对，有167个EST序列（占40．34％）与NCBI数据库中已知功能蛋白质具较高同源性，其中7个EST序列（占1．69％）与免疫防御相关基因同源；此外，204个EST（占49．28％）在数据库中未发现同源序列。研究表明，SSH技术能有效富集马氏珠母贝血淋巴差异表达基因，研究结果为探索马氏珠母贝免疫基因作用机理和调控机制奠定了基础。

外文摘要：The pearl oyster,Pinctada fucata,is the most important species cultured widely for production of marine pearls in the coastal provinces of South China, while mass mortalities of the cultured pearl oyster have occurred frequently in juvenile, mother and operated oysters. Understanding the immune defense mechanisms of the pearl oyster may contribute to the development of novel management strategies for diseases control and the long-term sustainability of pearl industry. In this study, a suppression subtractive hybridization （SSH） cDNA library was constructed from the hemocytes of P. fucata challenged with Vibrio alginolyticus, and fl- actin gene was used as internal control to estimate the efficiency of subtractive cDNA. A total of 600 clones in the SSH library were randomly selected and sequenced. The results showed that fl-actin was subtracted significantly at about 21^10 folds, suggested that the subtractive cDNA library was successfully constructed. PCR analysis showed that the inserts were 100 -750 bp in length. Based on BLAST searches in NCBI, 167 ESTs （40.34%） exhibited homology with genes of known functions, and 7 of the 167 ESTs （ 1. 69% ） were possibly related to immune and stress functions, and the remaining 204 ESTs had no significant matches with known sequences in the GenBank database. Altogether,the results obtained from this paper provide valuable information for further investigating the molecular immune mechanism of P. fucata.