[1]李 荔,李启沅,刘志刚,等.缢蛏过敏原的分离、鉴定与纯化[J].深圳大学学报理工版,2007,24(4):436-440.
 LI Li,LI Qi-yuan,LIU Zhi-gang,et al.Purification and identification of allergens in Razor clam[J].Journal of Shenzhen University Science and Engineering,2007,24(4):436-440.
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缢蛏过敏原的分离、鉴定与纯化()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第24卷
期数:
2007年4期
页码:
436-440
栏目:
生命科学
出版日期:
2007-10-30

文章信息/Info

Title:
Purification and identification of allergens in Razor clam
文章编号:
1000-2618(2007)04-0436-05
作者:
李 荔李启沅刘志刚余 晓
深圳大学生命科学学院,深圳518060
Author(s):
LI Li LI Qi-yuanLIU Zhi-gangand YU Xiao
College of Life Science,Shenzhen University,Shenzhen 518060,P.R.China
关键词:
缢蛏过敏原聚丙烯酰胺凝胶电泳免疫印迹高压液相色谱
Keywords:
Razor clamallergenSDS-PAGEWestern-blottinghigh pressure liquid chromatography
分类号:
R 392.8
文献标志码:
A
摘要:

以磷酸盐缓冲液提取蛋白,通过聚丙烯酰胺凝胶电泳(SDS-PAGE)对其抗原成分进行分析,选用缢蛏过敏患者的阳性血清进行免疫印迹,鉴定出主要与次要过敏原.用高压液相色谱(HPLC)纯化主要过敏原蛋白,鉴定其免疫活性,结果表明,缢蛏粗提液SDS-PAGE显示有18条蛋白条带,含量高的蛋白有6条,其分子量分别为110 kD、55 kD、42 kD、38 kD、35 kD和28 kD,其中以35 kD处最为富集.经Western-Blotting鉴定,58kD蛋白为缢蛏的主要过敏原,38 kD、28 kD和12 kD蛋白为缢蛏的次要过敏原.HPLC体积排阻纯化后得9个峰,以SDS-PAGE检测,58 kD的蛋白位于第4峰,其中有一条明显的蛋白条带.将该蛋白条带用HPLC反相纯化,得到两个峰.经Western-Blotting鉴定,58 kD的主要过敏原位于反相纯化的第2峰.

Abstract:

To purify and characterize the allergens of Razor clam(Sinonovacula constricta),the protein was extracted with phosphate buffer solution.The allergens in Razor clam were characterized with 9 allergic patients’ serum by Western-blotting after SDS-PAGE.To purify the primary allergen,HPLC method was employed.Finally immune activeness of the primary allergen was characterized by Western-blotting.The results indicate that,in the crude extracts,there are eighteen protein bands visible on the SDS-PAGE gel.The most abundant six bands are 110 kD,58 kD,42kD,38 kD,35 kD and 28 kD proteins,respectively.We identified the primary allergen whose molecular mass is 58 kD by Western-blotting,and the minor allergens whose molecular mass is 38 kD,28 kD and 12 kD,respectively.Upon HPLC-exclusion chromatogramphy of the crude extract,we obtained 9 peaks.After SDS-PAGE,the 58 kD protein was found in the forth peak.The protein in the forth peak was further purified by reversed phase high performance liquid chromatography.We obtained 2 peaks from the second purification.After Western-blotting ,we found that the 58 kD primary allergen was in the second peak.Thus,we obtained the pure primary allergen in Razor clam.

备注/Memo

备注/Memo:

收稿日期:2007-05-05;修回日期:2007-08-29
基金项目:广东省科技计划重点资助项目(2003A3080502);广州市科技计划重点资助项目(2002Z2-E4021);深圳市科技计划资助项目(200326)
作者简介:李荔(1968-),女(汉族), 辽宁省岫岩市人,深圳大学副教授、博士.E-mail:ylili@szu.edu.cn

更新日期/Last Update: 2007-12-05