作　　者： ; ; ; ; ;

机构地区： 湖南农业大学园艺园林学院

出　　处： 《湖南农业大学学报（自然科学版）》 2010年第3期300-303,共4页

摘　　要： 利用三氯乙酸-丙酮法、Tris-HCl提取法、改良PBS法提取柑橘叶片蛋白质,对提取蛋白质进行SDS-PAGE电泳、质谱鉴定和Western blot分析,结果表明,用改良PBS法提取的蛋白质质量浓度达30μg/μL,其SDS-PAGE电泳条带清晰,无杂质干扰,能达到质谱对样品的要求.Western blot分析结果表明,与抗柑橘溃疡病相关的PthA蛋白质在转pthA-nls基因糖橙和转pthA-nls基因冰糖橙中成功表达,pthA-nls基因已整合进甜橙基因组,表达蛋白质具有免疫反应活性. The Citrus were used to extract protein and optimized by the TCA-acedone,Tris-HCl extraction and improved method of PBS,and then after SDS-PAGE,mass spectrometry and Western blot.The result showed protein concentration is 30 μg/μL by the improved of PBS extraction methods.The SDS-PAGE result presented the bright and clear bands,with low background and could be identified by mass spectrometry.Furthemore,by applying transgenic pthA-nls gene in sweet orange plants,the PthA protein which is related to the Xanthomonas axonopodis pv.citri.could be found.To be concluded,the pthA-nls gene has been transferred into sweet orange and the expressive protein produced immunoreaction.

关 键 词： 柑橘叶片 蛋白质 溃疡病 免疫印记

领　　域： [生物学]