三孢布拉氏霉菌番茄红素环化酶多克隆抗体的制备及鉴定Preparation and Identification of Polyclone Antibody Against Blakeslea trispora CarRA
李晔,危晴,金丽华,王晓杰,辛秀兰,苏东海,袁其朋
摘要(Abstract)：
对三孢布拉氏霉菌番茄红素环化酶(CarRA)进行抗原表位分析并制备兔多克隆抗体。运用GOLDENKEY分子生物学软件对三孢布拉氏霉菌CarRA结构蛋白的抗原表位及其二级结构进行了分析比较,结合现有的抗原表位选取原则从中筛选了一段显示表位特征的氨基酸残基序列,应用固相合成法合成三孢布拉氏霉菌CarRA多肽,然后将其与KLH偶联制成免疫原,应用于动物免疫及多克隆抗体的制备,对该多抗与三孢布拉氏霉菌CarRA的反应性进行了检测。获得了三孢布拉氏霉菌CarRA的多抗,该抗体经间接酶联免疫吸附测定法检测其效价为1:32000,Dot blot检测在三个稀释度下都出现了目的条带,证实该抗体具有较好的反应性和特异性。成功制备了三孢布拉氏霉菌CarRA多抗,为进一步明确发酵促进剂的作用机制及其对三孢布拉氏霉菌代谢网络的影响,对CarRA进行定量以及对其相关功能进行深入研究提供了有利条件。
关键词(KeyWords)： 三孢布拉氏霉菌;CarRA;多肽合成;多克隆抗体;制备;鉴定
基金项目(Foundation): 国家自然科学基金(21176018,21306002);; 北京电子科技职业学院院内重点课题(YZKB2013009);; 北京高等学校“青年英才计划“(YETP1801);; 科研基地建设——科技创新平台项目——酶制剂研发基地建设项目PXM2014_014306_000064
作者(Author): 李晔,危晴,金丽华,王晓杰,辛秀兰,苏东海,袁其朋
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