青岛浒苔完整叶绿体分离及其蛋白质提取方法的初步建立  

Preliminary establishment of extraction methods for intact chloroplasts and its protein components from Ulva prolifera

作　　者：杜智欣[1,2] 邱艳红[1,2] 鲁洁[2] 段维军[3] 孔君[2] 朱水芳[2] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193 [2]中国检验检疫科学研究院植物检疫研究所 [3]宁波出入境检验检疫局

出　　处：《植物检疫》2013年第2期44-48,共5页Plant Quarantine

基　　金：中国检科院专项(2009 JK002)

摘　　要：初步建立青岛海域漂浮浒苔完整叶绿体的分离纯化及其叶绿体蛋白质提取方法,为进一步研究浒苔叶绿体蛋白质组学奠定基础。本研究应用组织研磨结合Percoll密度梯度离心的方法分离出浒苔的完整叶绿体,随后以两种蛋白裂解液配方分别提取叶绿体总蛋白,并通过蛋白定量和两种聚丙烯酰胺凝胶电泳进行比较分析。结果表明:Percoll密度梯度离心的方法能够获得完整且较纯的浒苔细胞叶绿体;含有尿素以及CHAPS的蛋白裂解液抽提的叶绿体蛋白质数量比SDS裂解液更多;相较于普通Tris-SDS-PAGE,采用Tricine-SDS-PAGE电泳系统可以有效分离40kDa以下的小分子叶绿体蛋白质。In order to study the chloroplast proteomics of Ulva prolifera, the extraction methods for intact chloroplast and its protein components from Ulva prolifera collected from the surface of Qingdao sea area were preliminarily established. In this study, the samples of Ulva prolifera were broken using tissue grinding and the intact chloroplasts were isolated by centrifugation with Percoll gradient. Then, two kinds of lysis buffer for protein extractionwere utilized to extract the protein components of the chloroplasts and the extraction efficiencies of the buffers were compared by concentration determination and two different methods of SDS - PAGE gel electrophoresis. The results indicated that Percoll density gradient centrifugation methods could obtain intact chloroplasts in high purity ; the protein components obtained by lysis buffer containing Urea and CHAPS were more than those obtained by lysis buffer containing SDS. In addition, compared with Tris - SDS - PAGE, the proteins of less than 40kDa from intact chloroplasts were more effectively separated by Tricine -SDS -PAGE.

关 键 词：浒苔 叶绿体 蛋白质 密度梯度离心 提取 

分 类 号：Q946[生物学—植物学]