宝石鲈肝胰脏蛋白质双向电泳方法的摸索与优化

天津农学院学报 ›› 2013, Vol. 20 ›› Issue (2): 15-17.

宝石鲈肝胰脏蛋白质双向电泳方法的摸索与优化

周鹏¹, 宋立伟¹, 白东清¹, 孙向军², 齐野¹, 乔秀亭^{1,通信作者}

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

收稿日期:2013-01-05 发布日期:2019-10-18
通讯作者: 乔秀亭（1965-）,男,天津市人,教授,硕士,主要从事水产营养与饲料方面的研究。E-mail：qxt6510@yahoo.com.cn。
作者简介:周鹏（1987-）,男,天津市人,硕士在读,主要从事水产营养与饲料方面的研究。E-mail：305179365@qq.com。
基金资助:
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）

Establishment and Optimization of 2-DE Techniques in Hepatopancreas Proteome of Jade perch

ZHOU Peng¹, SONG Li-wei¹, BAI Dong-qing¹, SUN Xiang-jun², QI Ye¹, QIAO Xiu-ting^{1,Corresponding Author}

1. Department of Fishery Science, Tianjin Agricultural University, Tianjin 300384, China;
2. Beijing Fisheries Research Institute, Beijing 100068, China

Received:2013-01-05 Published:2019-10-18

摘要/Abstract

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 mM Tris-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,10 000 V·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。

关键词: 宝石鲈, 肝胰脏, 蛋白质组, 双向电泳

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 І（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.

Key words: Jade perch（Scortum Barcoo）, hepatopancreas, proteome, 2-DE

中图分类号:

S965.211

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, 20(2): 15-17.

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 University, 2013, 20(2): 15-17.

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参考文献

[1] Pennington S R,Wilkins M R,Hochstrasser D F,et al.Proteome analysis:From protein characterization to biological function[J]. Trends in Cell Biology,1997,7(4):168-173.
[2] Stanley B A,Neverova I,Brown H A,et al.Optimizing protein solubility for two-dimensional gel electrophoresis analysis of human myocardium. Proteomics[J]. J Biol Chem Biophys Methods,2003,3(6):815-820.
[3] O’ Farrell P H. High resolution two-dimensional eletrophoresis of proteins[J]. J Biol Chem,1975,250(10): 4007-4021.
[4] Mezhoud K,Bauchet A L,Chateau-Joubert S,et al.Proteomic and phosphoproteomic analysis of cellular responses in medaka fish(Oryzias latipes)following oral gavage with microcystin-LR[J]. Toxicon,2008,51(8):1431-1439.
[5] 吴海庆,李明云,叶帅东,等. 大弹涂鱼肝脏蛋白质双向电泳技术的建立及优化[J]. 生命科学仪器,2009,8(7):29-32.
[6] Keyvanshokooh S,Vaziri B.Proteome analysis of Persian sturgeon(Acipenser persicus)ova[J]. Animal Reproduction Science,2008,109(1-4):287-297.
[7] Keyvanshokooh S,Kalbassi M R,Hosseinkhani S,et al.Comparative proteomics analysis of male and female Persian sturgeon(Acipenser persicus) gonads[J]. Animal Reproduction Science,2009,111(2-4):361-368.
[8] Wu Y J,Wang S Y,Peng X X.Serum acute phase response(APR)-related proteome of loach to trauma[J]. Fish & Shellfish Immunology,2004,16(3):381-389.
[9] Tang J Z,Zhang D Y,Cheng J,et al.Comparative analysis of theamino acid composition and proteomic patterns of the muscle proteins from two teleosts,Siniperca chuatsi L. and Hypophth-almichthys molitrix L.[J]. Journal of Fisheries of China,2007,31(3):361-368.
[10] K ruger N J. The Bradford method for protein quantitation[J]. Methods Mol Biol,1994,32(1):9-15.
[11] Grog A,Postel W,Günther S.The current state of two-dimensional electrophoresis with immobilized pH gradients[J]. Electrophoresis,2002,21(6):1037-1053.
[12] 郭尧君. 蛋白质电泳实验技术[M]. 2版. 北京:科学出版社,2005:213-214.