渔业科学进展2011，Vol.32Issue(2)：41-46,6.
大菱鲆表皮蛋白质组双向电泳技术的建立及优化
Establishment and optimization of two-dimensional gel electrophoresis for epidermal proteome of turbot Scophthalmus maximus
摘要
Abstract
The two-dimensional gel electrophoresis for the epidermal proteome of Scophthalmus maximus was established and optimized through comparative tests by using different extraction methods, isoelectric focusing programs and sample volumes.The total proteins of the turbot epidermis were extracted by the combination of improved trichloroacetic acid(TCA)/ acetone and ultrasonic extractions.The lysis buffer based on the Bio-Rad recommendations was optimized.Protein of 250 μg was loaded on the 17cm pH 4-7 immobilized pH gradients (IPG) strips.The programs for the gel isoelectric focusing were 30V linear ramp, 6h/250V linear ramp, 2.5h/500V linear ramp, 1.5h/1000V rapid, 2h/4000V linear ramp, 3h/8000V rapid,3h/8000V linear ramp, and 1h/500V 24h.The gel was stained with sliver nitrate, scanned and analyzed using the Image Master 2D Platinum 7.0 software.The two-dimensional gel electrophoresis analysis of the epidermal proteome of the turbot was successfully established with high quality map of separation under the optimization of the experimental conditions, which will be a valuable preparation for further epidermal proteomics research of S.maximus.关键词
大菱鲆/表皮组织/蛋白质/双向电泳Key words
Turbot Scophthalmus maximus/ Epidermis / Proteome/ 2-DE分类
生物科学引用本文复制引用
薛宝贵,黄智慧,周洲,马爱军..大菱鲆表皮蛋白质组双向电泳技术的建立及优化[J].渔业科学进展,2011,32(2):41-46,6.基金项目
现代农业产业技术体系建设专项资金(nycytx-50)、国家支撑计划专题(2006BAD01A12012)、农业公益性行业科研专项经费项目(nyhyzx07046)和国家863计划课题(2006AA100302)共同资助 （nycytx-50）
