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.
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)
关键词
大弹涂鱼
肝脏
蛋白质组
双向电泳
建立
优化
Boleophthalmus pectinirostnx,, liver
, proteome
2-DE
establishment
Optimization
