| 引用本文: | 沈 妍,刘 伟,湛垚垚,黄显雅,穆琳琳,徐 蕾,丁 君,常亚青.虾夷扇贝性腺和闭壳肌蛋白质双向电泳分析[J].海洋科学,2014,38(6):10-16. |
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| 虾夷扇贝性腺和闭壳肌蛋白质双向电泳分析 |
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沈 妍1,2, 刘 伟1,2, 湛垚垚1,2, 黄显雅1,2, 穆琳琳1, 徐 蕾1, 丁 君1,2, 常亚青1,2
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1.大连海洋大学 水产与生命学院;2.农业部北方海水增养殖重点实验室
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| 摘要: |
| 本研究建立了虾夷扇贝性腺、闭壳肌蛋白质双向电泳体系, 比较分析了不同等电聚焦时间对虾夷扇贝性腺、闭壳肌蛋白质双向电泳结果的影响, 进一步利用该双向电泳系统分离了性腺和闭壳肌可溶性全蛋白, 并利用计算机图像分析软件对虾夷扇贝性腺和闭壳肌蛋白质双向电泳图谱进行了检测分析。实验结果表明, 采用细胞超声破碎-丙酮沉淀除盐–50 mmol/L Tris-HCl (pH6.8)复溶, 1000V, 3 h 等电聚焦, 12%SDS-PAGE 和高灵敏度银氨染色, 可获得分辨率较高、重复性良好的双向电泳图谱。通过PDQuest8.0 图像分析软件检测分析, 发现pH5～8 条件下, 虾夷扇贝性腺和闭壳肌组织的蛋白质主要分布于pH5～7, 相对分子质量97～20.1 ku 范围, 性腺组织双向电泳图谱检出的蛋白质斑点数为265 个, 闭壳肌组织检出161 个蛋白质斑点。结果显示, 虾夷扇贝性腺、闭壳肌可溶性蛋白质表达谱各具特点, 性腺组织可溶性蛋白种类较闭壳肌组织丰富。本研究实验结果可为进一步研究扇贝蛋白质组学以及功能基因组学奠定一定的基础。 |
| 关键词: 虾夷扇贝 性腺 闭壳肌 双向电泳 电泳图谱分析 |
| DOI：10.11759/hykx20120910001 |
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| 基金项目:海洋公益项目(2012 05031); 国家“863”计划项目(2012AA10AA410);国家“948” 项目(2011 12G); 校博士启动基金项目(SYYJ2010002) |
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| Establishment of two-dimensional electrophoresis system and analysis of two-dimensional electrophoresis map for gonad and muscle of scallop Patinopecten yessoensis |
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| Abstract: |
| The carrier ampholytes pH gradient two-dimensional electrophoresis for proteins of gonad and muscle tissues of scallop Patinopecten yessoensis was established and optimized. The protein expression of gonad and muscle tissues was investigated by the established two-dimensional electrophoresis system with computer image analysis software. The total proteins of gonad and muscle were extracted with method of ultrasonication-acetone sedimentate-50 mmol/L Tris-HCl (pH 6.8), and separated by 1000 volt 3-hour isoelectrofocusing and 12% SDS-PAGE. The maps of two-dimensional electrophoresis of gonad and muscle proteins were analyzed by PDQuest 8.0 image analysis software. Results showed that gonad and muscle proteins were distributed mainly in pH range of 5～7 and molecular weight range of 97～20.1 ku. In pH 5～8, gonad and muscle proteins contained 265 and 161 protein spots, respectively. The results suggested that there is a significant tissue specific difference at protein level between gonad and muscle of scallop P. yessoensis, which might be useful for further proteomics and functional genomics studies on scallop P. yessoensis. |
| Key words: Patinopecten yessoensis gonad muscle two-dimensional electrophoresis two-dimensional electrophoresis map analysis |