学术
摘 要:
目的分析并用免疫和质谱方法鉴定枝孢芽枝菌（Cladosporium cladosporioides）的主要过敏原。方法空气中暴皿获得枝孢芽枝菌菌种,经鉴定后大规模培养;用碳酸盐缓冲液提取枝孢芽枝菌蛋白,并用SDS-PAGE、Western-blotting方法鉴定出枝孢芽枝菌的主要过敏原。主要过敏原胶内酶切,再经基质辅助激光解吸电离飞行时间质谱（MALDI-TOF-MS）分析,所得质谱数据进入网站（http：//www.matrixscience.com）Mascot检索比对。结果经过暴皿获得枝孢芽枝菌菌种,其蛋白粗提液SDS-PAGE结果显示有14个条带,含量丰富的有10个条带;Western-Blotting鉴定出18kD的蛋白阳性率达66%,为枝孢芽枝菌的主要过敏原;质谱显示枝孢芽枝菌主要过敏原与约18kD嗜冷杆菌（Psychrobacter arcticum）50S的核糖体蛋白的匹配分值最高。结论采用免疫、质谱法鉴定了枝孢芽枝菌的主要过敏原是约18kD的核糖体蛋白。[著者文摘]
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文章出处:
《卫生研究》-2008年37卷1期 -50-52页
栏目信息:
文献标识码:
A
文章编号:
1000-8020（2008）01-0050-03
Immunological analysis and mass spectrometry identification of the major allergen from Cladosporium cladosporioides
LI Li, LI Qiyuan, LIU Zhigang, WU Haiqiang （Allergy and Immunology Institute, Shenzhen University, Shenzhen 518060, China）
Abstract:
Objective To analyze and identify the major allergen from Cladosporium cladosporioides. Methods The Cladosporium cladosporioides were collected by means of glass dish exposed by air. After they were cultured, the protein of the Cladosporium cladosporioides was extracted with carbonate buffer solution. The major allergen in the Cladosporium cladosporioides was characterized by Western-blotting after SDS-PAGE. After in-gel digestion, the peptide mass values of the major allergen which were obtained by matrix-assisted laxer desorption ionization time-of-night mass spectrometry, （MALDI-TOF-MS） were used as inputs to search corresponding proteins with the Mascot （http：//www. matrixscience. corn）. Results There were fourteen protein bands visible on the SDS-PAGE gel in the crude extracts. The most abundant ten bands were 72kD, 61kD, 42kD, 36kD, 35kD, 34kD, 24kD, 22kD, 18kD and 14kD proteins, respectively. The protein whose molecular mass was 18kD was identified by Western-blotting as major allergen. The major allergen was similar to the 18kD ribosomal protein of Psychrobacter arcticum based on peptide mass fingerprint analysis （MALDI-TOF- MS）. Conclusion The major allergen from Cladosporium cladosporioides, a ribosomal protein about 18kD, was analyzed and identified.[著者文摘]
Key words:
Cladosporium cladosporioides, allergen, Western-blotting, mass specrometry, identification
基金资助:
广东省科技重点专项（No.2003A3080502）;广东省自然科学基金项目（No.040636）;深圳市科技计划资助项目（No.200326）
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