应用羟基磷灰石层析分离纯化鼠疫菌Pla蛋白的研究  

Study on separation and purification of Pla protein of Yersinia pestis by hydroxyapatite chromatography

作　　者：杜春红[1] 唐雪[1] 石丽媛[1] 王鹏[1] 杨光璨[1] 董珊珊[1] 宋志忠[1] 

机构地区：[1]云南省地方病防治所云南省鼠疫防控技术重点实验室,云南大理671000

出　　处：《疾病预防控制通报》2011年第5期10-12,共3页Bulletin of Disease Control & Prevention(China)

基　　金：云南省科技攻关项目(2007CA010)

摘　　要：目的应用CHT陶瓷羟基磷灰石层析柱分离纯化鼠疫耶尔森氏菌纤溶酶原激活因子(Pla)。方法优化层析条件,对采用超声破碎结合硫酸铵盐析得到的Pla粗提样品进行纯化,收集纯化中各洗脱峰样品,进行SDS-PAGE。结果上样缓冲液中添加钙离子,可促进Pla与层析柱的结合,经纯化后的Pla,去除了大部分杂蛋白,得到主要由相对分子质量约为31×103、35×103、37×103 Da和3条蛋白带为主的Pla,经软件分析计算,占蛋白总量约80%。结论 CHT陶瓷羟基磷灰石层析能实现对Pla的基本纯化。Objective To separate and purify plasminogen activator（Pla） from Yersinia pestis by ceramic hydroxyapatite（CHT） chromatography.Methods Crude Pla collected by sonication and precipitation with ammonium sulfate was purified by optimized chromatographic conditions,samples collected in the elution peaks were collected and analyzed by SDS-PAGE.Results Adding calcium ions to sample buffer enhanced target proteins＇ ability to bind to the column.Proteins with molecular mass of approximately 31×103,35×103 and 37×103 Da which accounted for about 80% of total proteins were obtained by purification.Conclusions The purified Pla can be obtained by ceramic hydroxyapatite chromatography.

关 键 词：羟基磷灰石层析 鼠疫 Pla蛋白 分离纯化 

分 类 号：R378.61[医药卫生—病原生物学]