| 宝石鲈肝胰脏蛋白质双向电泳方法的摸索与优化 |
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| 引用本文： | 周鹏,宋立伟,白东清,孙向军,齐野,乔秀亭.宝石鲈肝胰脏蛋白质双向电泳方法的摸索与优化[J].天津农学院学报,2013(2):15-17,21. |
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| 作者姓名： | 周鹏 宋立伟 白东清 孙向军 齐野 乔秀亭 |
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| 作者单位： | [1]天津农学院水产科学系,天津300384 [2]北京市水产科学研究所,北京100068 |
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| 基金项目： | 天津市科委科技攻关项目“宝石鲈、香鱼集约化高效养殖技术研究”（043121211）;北京市科学技术委员会科技计划项目“北京水产品种更新关键技术研究与应用”（Z080005032508019） |
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| 摘 要： | 选取宝石鲈肝胰脏作为样品,通过优化裂解配方及摸索裂解液与样品的最适比例,设计两个等电聚焦程序,建立和优化宝石鲈鱼肝胰脏蛋白质双向电泳实验方法,为宝石鲈肝胰脏内目标蛋白的研究提供参考。结果表明,采用裂解液配方:8 M Urea,2 M Thiourea,0.5%(W:V)CHAPS,40 mMTris-Base,1%DTT,2%(W:V)Pharmalyte pH 3～10,1 mM PMSF;裂解液和样品液比为1∶3时裂解效果较好;等电聚焦程序I(S1:0～500 V,500 V.h;S2∶500 V,500 V.h;S3∶500～3 500 V,10 000V.h;S4∶3 500 V,50 000 V.h,S5∶3 500～5 000 V,8 000 V.h)具有最好的等电聚焦效果。
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| 关 键 词： | 宝石鲈 肝胰脏 蛋白质组 双向电泳 |
Establishment and Optimization of 2-DE Techniques in Hepatopancreas Proteome of Jade perch |
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| ZHOU Peng,SONG Li-wei,BAI Dong--qing,SUN Xiang-jun,QI Ye,QIAO Xiu-ting.Establishment and Optimization of 2-DE Techniques in Hepatopancreas Proteome of Jade perch[J].Journal of Tianjin Agricultural College,2013(2):15-17,21. |
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| Authors: | ZHOU Peng SONG Li-wei BAI Dong--qing SUN Xiang-jun QI Ye QIAO Xiu-ting |
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| Institution: | 1 Corresponding Author ( 1. Department of Fishery Science, Tianjin Agricultural University, Tianjin 300384, China; 2. Beijing Fisheries Research Institute, Beijing 100068, China) |
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| Abstract: | Using the hepatopancreas of Jade perch as the experimental samples, the tissue lysate formula and ratio of lysis buffer volume to tissue weight, as well as the isoelectric focusing electrophoresis procedures were screened. The results were as follows： lysate formula I （8 M Urea, 2 M Thiourea, 0.5% （ W ; V） of CHAPS, 40 mM Tris-Base, 1% DTT, 2% （ W ; V） Pharmalyte pH 3-10, 1 mM PMSF） was the better. When the ratio Lysate volume to the sample weight was 1 ： 3, the lytic effect was the better. The electrophoretic program I （S1 ; 0-500 V, 500 V.h, S2 ; 500 V, 500 V.h, S3 ; 500-3 500 V, 10 000 V.h, S4 ; 3 500 V, 50 000 V.h. S5 ; 3 500-5 000 V, 8 000V.h） had the better isoelectric focused effect. Based on these results, a protocol of two-dimensional gel electrophoresis （2-DE） was set up. The optimized 2-DE techniques provided a basis for studying the proteome of Jade perch hepatopancreas. |
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| Keywords: | Jade perch （Scortum Barcoo） hepatopancreas proteome 2-DE |
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