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棉花学报  2013 

棉花纤维素合酶复合体蛋白的分离与鉴定

DOI: 1002-7807(2013)02-0129-06, PP. 129-134

Keywords: 棉纤维,纤维素合酶,复合体,次生壁

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Abstract:

选用开花后24d的棉纤维提取原生质膜,用TritonX-100溶解后通过纤维素合酶1(Gossypiumhirsutumcellulosesynthase1,GhCESA1)抗体进行免疫共沉淀(Co-Immunoprecipitation,Co-IP),以聚丙烯酰胺凝胶电泳(Sodiumdodecylsulfatepolyacrylamidegelelectrophoresis,SDS-PAGE)分离免疫共沉淀的产物,用液相色谱-质谱联用仪(Liquidchromatography-massspectrometry,LC-MS)进行鉴定。结果表明,棉纤维纤维素合酶复合体可能存在68种蛋白,其中包括8种纤维素合酶(Cellolosesynthse,CESA),它们涵盖了棉纤维初生壁和次生壁2种类型纤维素合酶。这说明在次生壁形成的细胞中存在2种类型CESA。本研究也表明复合体中存在非CESA蛋白。

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