柑橘叶片蛋白质高效提取与溃疡病PthA蛋白质Western blot分析  

Extraction of Citrus leaf protein and analysis of Western blot of the Citrus canker protein PthA

作　　者：李娜[1,2] 覃磊[1] 严佳文[1] 陈佩[1] 邓子牛[1,2] 

机构地区：[1]湖南农业大学园艺园林学院,湖南长沙410128 [2]湖南省作物种质创新与资源利用重点实验室,湖南长沙410128

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Journal of Hunan Agricultural University(Natural Sciences)

基　　金：国家自然科学基金项目(30871702);湖南省科学技术厅项目(2008WK2007)

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 

分 类 号：Q946.1[生物学—植物学]