大弹涂鱼肝脏蛋白质双向电泳技术的建立及优化  

Establishment and Optimization of 2-DE Technique in Liver Proteome of Boleophthalmus pectinirostris

作　　者：吴海庆[1] 李明云[1] 叶帅东[1] 冀德伟[1] 

机构地区：[1]宁波大学生命科学与生物工程学院,浙江宁波315211

出　　处：《生命科学仪器》2009年第8期33-36,共4页Life Science Instruments

基　　金：长江学者和创新团队发展计划(IRT0734);浙江省财政厅和浙江省海洋与渔业局资助项目(503050005)

摘　　要：通过对不同裂解液配方、等电聚焦程序和SDS-PAGE方式等的对比实验,建立和优化了大弹涂鱼肝脏蛋白质组双向电泳的相关技术体系。结果显示采用裂解液中添加Tris和TBP,聚焦时适当延长除盐时间,提高聚焦电压和功率(伏-小时),能显著提高双向电泳图谱的分辨率,MSOG程序进行双向电泳(Multi-strips on One Gel MSOG),提高了双向凝胶的匹配率及有效性,降低了人为修饰点的增加,匹配率高达90%。通过相关条件优化提高了大弹涂鱼肝脏蛋白双向电泳图谱的分辨率,为后续大弹涂鱼的毒理蛋白质组学研究提供技术保障。In this paper, the 2-DE related techniques in liver proteome of Boleophthalmus pectinirostris was constructed and optimized by comparative tests between different extraction methods, isoelectric focusing Programs and the way of SDS-PAGE. The results showed that： It could significantly increase the resolution of 2-DE profiles after adding Tris and TBP in lysis buffer, prolonging the time of desalting and increasing the Voltage and power （Volt-hours） of isoeleetrie focusing properly, SDS-PAGE in Multi-strips on One Gel matching rate reached 90% With optimization of the experimental conditions, 2-DE in liver proteome of Boleophthalmus pectinirostris has been successfully established, which could provide technical support for follow-up toxicological proteomics research of Boleophthalrnus pectinirosttis.

关 键 词：大弹涂鱼 肝脏 蛋白质组 双向电泳 建立 优化 

分 类 号：Q510.3[生物学—生物化学] Q51