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口虾蛄性腺组织蛋白质双向电泳体系的建立及优化
引用本文:莫海波,田燚,湛垚垚,赵丽娜,刘海映.口虾蛄性腺组织蛋白质双向电泳体系的建立及优化[J].生物技术通报,2012(4):152-157.
作者姓名:莫海波  田燚  湛垚垚  赵丽娜  刘海映
作者单位:大连海洋大学农业部北方海水增养殖重点开放实验室,大连,116023
基金项目:国家自然科学基金项目,海洋公益性行业科研专项
摘    要:旨在通过口虾蛄雄性、雌性性腺组织蛋白质双向电泳技术体系的优化,获得雄性、雌性口虾蛄性腺蛋白质的表达图谱。结果表明,不同的蛋白提取方法,上样前处理方法,上样量,聚焦时间及雌、雄性口虾蛄性腺蛋白表达图谱存在一定的差异。雌性、雄性口虾蛄用Tris-HCl提取后用丙酮沉淀方法提取蛋白质、不经上样缓冲液处理、上样量为10μg时,得到较好图谱。对图谱分析发现在pH4-6.5范围内,雌性口虾蛄性腺可溶性蛋白质种类多于雄性。雄性口虾蛄蛋白质点相对于雌性较少,且蛋白质大部分分布于酸性端。经过蛋白质双向电泳体系的优化,能显著提高双向电泳图谱的分辨率,为进一步研究口虾蛄性别差异表达蛋白的筛选,后续的口虾蛄蛋白质组学研究提供技术保障。

关 键 词:口虾蛄  性腺蛋白质  双向电泳

Preliminary Establishment of Two-dimensional Gel Electrophoresis for Gonad Tissue of Oratosquilla oratoria
Mo Haibo , Tian Yi , Zhan Yaoyao , Zhao Lina , Liu Haiying.Preliminary Establishment of Two-dimensional Gel Electrophoresis for Gonad Tissue of Oratosquilla oratoria[J].Biotechnology Bulletin,2012(4):152-157.
Authors:Mo Haibo  Tian Yi  Zhan Yaoyao  Zhao Lina  Liu Haiying
Institution:Mo Haibo Tian Yi Zhan Yaoyao Zhao Lina Liu Haiying(Key Laboratory of Mariculture,Ministry of Agriculture,Dalian Ocean University,Dalian 116023)
Abstract:The gonad two-dimensional electmphoresis system in Oratosquilla oratoria was optimized and carried out.The purpose of this research was to establish a suitable system for edible mantis shrimp to analyze proteomics.Several methods of protein preparation(TCA-acetone precipitation treatment,Tris-HCL treatment,Tris-HCL acetone treatment and pure water treatment)were applied to extract total protein from edible mantis shrimp gonad.The subsequent 2-DE analysis was carried out in order to obtain the best conditions.Experiments were optimized in protein extraction method,sampling methods,sample volume,focusing condition optimization.The results showed that different patterns were obtained with the different methods of protein extraction,sample pretreatment methods,sample volume,focus time in male and female gonad of edible mantis shrimp.The results indicated that protein preparation with the Tris-HCL acetone extraction treatment had the best effective with the most electrophoretic spots,and all the spots were evidently observed in female and male edible mantis shrimp.The 2-DE electrophoretic analysis in gonad get a satisfactory map using sample without buffer solution treatment,the sample size 10 μg.The results showed the spots of female gonadal were found in pH4-6.5 range,and more than male.
Keywords:Edible mantis shrimp Gonad protein Two-dimensional electrophoresis(2-DE)
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