大弹涂鱼肝脏蛋白质双向电泳技术的建立及优化

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。

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

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.