大黄鱼肝脏蛋白质组双向电泳技术的建立及优化  

Establishment and Optimization of 2-DE Technique in Liver Proteome of Large Yellow Croaker Pseudosciaena crocea

作　　者：李明云[1] 冀德伟[1] 吴海庆[1] 陈炯[1] 史雨红[1] 

机构地区：[1]应用海洋生物技术教育部重点实验室,宁波大学,浙江宁波315211

出　　处：《水产科学》2010年第1期27-30,共4页Fisheries Science

基　　金：国家高技术研究发展计划(“863”)项目(2006AA10A405);浙江省重大项目(2006C12005);宁波市重大项目(2008C10022);长江学者和创新团队发展计划项目(IRT0734)

摘　　要：通过对不同裂解液配方、等电聚焦程序和上样量等条件的对比试验,建立和优化了大黄鱼肝脏蛋白质组双向电泳的相关技术体系。试验结果表明,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,能显著提高双向电泳图谱的分辨率,而17 cm pH 5-8 IPG胶条的最佳上样量为2.0 mg。通过相关条件优化提高了大黄鱼肝脏蛋白双向电泳图谱的分辨率,为大黄鱼肝脏蛋白质组学的进一步研究奠定了基础。The 2-DE related techniques was constructed and optimized in liver proteome of large yellow croaker Pseudosciaena crocea by comparative tests between different extraction methods,isoelectric focusing programs and sample volume.The results showed that the resolution of 2-DE profiles was significantly increased by adding Tris and TBP in lysis buffer,prolonging the time of desalting and increasing the voltage and power（Volt-hours）of isoelectric focusing properly.The optimal sample volume of 17 cm pH 5~ 8 IPG strips was found to be 2.0 mg.The 2-DE in liver proteome of the fish was successfully established with high quality map of separation under the optimization of the experimental conditions,which stands as a valuable resource for further liver proteomics research of Pseudosciaena crocea.

关 键 词：大黄鱼 肝脏 蛋白质组 双向电泳 

分 类 号：S965.322[农业科学—水产养殖]