棉花纤维素合酶复合体蛋白分离与鉴定.pdfVIP

花 学 报 Cotton Science 20 13 25 渊2 冤院129~ 134 花纤维素合酶复合体蛋白的分离与鉴定 1,2 1 1 1* 先良 李傲 彭良才 夏 涛 1. 中农业大学作物遗传改良国家重点实验室/ 生物质与生物能源研究中心/ 植物科学技术学院/ 生命科学技术学院袁 武汉430070 曰2.荆楚理工学院生物工程学院袁湖北荆门448000冤 摘要院选用开花后24 d 的棉纤维提取原生质膜袁用Triton X- 100 溶解后通过纤维素合酶1 cellulose synthase 1, GhCESA1 冤抗体进行免疫共沉淀 Co-Immunoprecipitation, Co-IP 冤袁以聚丙烯酰胺凝胶电泳 Sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS冤分离免疫共沉淀的产物袁用液相色谱- 质谱联用仪 Liquid chromatography-mass spectrometry, LC-MS冤进行鉴定遥结果表 袁棉纤维纤维素合酶复合体 可能存在68 种蛋白袁其中包括8 种纤维素合酶 Cellolose synthse, CESA冤袁它们涵盖了棉纤维初生壁和次生壁 2 种类型纤维素合酶遥 这说明在次生壁形成的细胞中存在2 种类型CESA遥 本研究也表明复合体中存在非 CESA 蛋白遥 关键词院棉纤维曰纤维素合酶曰复合体曰次生壁 中图分类号院S562:Q814 文献标志码院A 文章编号院1002-7807 渊20 13冤02-0129-06 1,2 1 1 1* LI Xian-liang , LI Ao , PENG Liang-cai , XIA Tao (1. 430070, ; 2. 448000, ) To identify the components of CSCs, plasma membrane (PM) was extracted from cotton fibers at 24 days post anthesis (DPA) and solubilized by triton X-100, from which CSCs were isolated by co-immunoprecipitation mediated by cellulose synthase1(GhCESA 1). The proteins isolated by co-immunoprecipitation mediated by GhCESA 1 were character- ized by western-blotting using the antibodies of GhCESA 1 and sucrose synthase (GhSuSy). The results show there are GhCESA 1 and GhSuSy proteins, which indicates this method can isolate the components of CSCs. Then, CSCs proteins through SDSwere subj ected to nanoflow liquid chromatography-tandem mass spectrometry (LC-MS). 68 kinds of proteins were identificated, in which there were 8 kinds of CESA proteins. GhCESA3, 5, 6