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刺参(Apostichopus japonicus)腐皮综合症发生相关蛋白的分离与鉴定
张 鹏1, 李成华1, 李 晔1, 苏秀榕1, 金春华1, 李太武2
1.宁波大学海洋学院;2.宁波城市职业技术学院
摘要:
采用比较蛋白质组学的方法, 对2011年10月20日取自辽宁省普兰店室内养殖厂腐皮综合症刺参和健康个体肠组织中差异表达蛋白进行了研究。结果表明, 在优化的电泳条件(80μg脱盐蛋白、pH 4—7 [NL,13cm]的IPG预制胶条)下, 共检测到约700个蛋白质点; 表达量差异两倍以上的蛋白点51 个, 最大表达差异比率为4.32; 对其中重复性一致的30个差异点进行质谱鉴定, 成功鉴定了23个差异蛋白, 包括铁蛋白, 过氧化氢酶, 泛素相关修饰子3, 细胞色素氧化酶和肌动蛋白等; 进一步扫描实验室完成的刺参转录组数据, 筛选了过氧化氢酶和泛素相关修饰子3的EST序列。本研究是蛋白层面上刺参腐皮综合症发生机制的有效尝试, 研究结果为进一步认识该疾病的发生机制提供了参考。
关键词:  刺参  腐皮综合症  双向电泳  转录组
DOI:10.11693/hyhz201303031031
分类号:
基金项目:国家自然科学基金项目资助, 31101919 号, 41276120 号; 宁波市科技计划项目, 2011C11001 号; 宁波市农业攻关项目, 201301C1011001号; 浙江省人事厅151人才工程项目, 2011—2013; 宁波大学2012年度研究生科研创新基金资助项目, 2012—2013
ISOLATION AND CHARACTERIZATION OF DIFFERENTIAL EXPRESSED PROTEINS FROM SKIN ULCERATION SYNDROME APOSTICHOPUS JAPONICUS
ZHANG Peng1, LI Cheng-Hua1, LI Ye1, SU Xiu-Rong1, JIN Chun-Hua1, LI Tai-Wu2
1.School of Marine Sciences, Ningbo University;2.Ningbo City College of Vocational Technology
Abstract:
The differential expressed proteins between healthy and skin ulceration syndrome Apostichopus japonicus collected from indoor ponds of Pulandian Hatchery in Dalian, China in October 20, 2011 were investigated by comparative proteomics in the present study. All the results showed: (1) total of 700 spots were successfully identified under optimal conditions (80?g desalted protein and pH 4—7 IPG strip [NL, 13cm]); (2) 51 spots showed significant differences in abundance with the maximum difference ratio of 4.32; (3) 30 spots with the higher reproducibility were characterized by MS, among which 23 spots including ferritin, catalase, ubiquitin-related modifier 3, cytochrome oxidase, and actin were identified from protein database; (4) the partial cDNA sequences of catalase and ubiquitin-related modifier 3 were obtained by screening our transcriptome library. This study is the first analysis of differentially expressed proteins in the diseased sea cucumber at the protein level and provides a new insight for further study in skin ulceration syndrome outbreak in the species.
Key words:  Apostichopus japonicus  skin ulceration syndrome  2-Dimensional electrophoresis  transcriptome
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