重组血红素加氧酶-2在大肠杆菌中表达与纯化方法的研究  

Expression and purification for heme oxygenase-2 recombinant in E.coli BL-21

作　　者：滕悦秋[1] 王秀宏[1] 赵炜明[1] 王志刚[1] 刘明[1] 周虹[1] 

机构地区：[1]哈尔滨医科大学生物化学与分子生物学教研室,黑龙江哈尔滨150086

出　　处：《哈尔滨医科大学学报》2004年第1期1-4,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目 ( 3 0 0 0 0 0 3 0 ) ;黑龙江省自然科学基金资助项目 (D0 0 0 4)

摘　　要：目的　确定重组血红素加氧酶 2在大肠杆菌中表达与纯化方法。方法　将已构建的血红素加氧酶 2(HO 2 )的重组质粒pMW1 72a HO 2转入大肠杆菌BL 2 1 ,分析不同温度及摇床转速对HO 2表达的影响 ,确定可溶性表达最佳条件。使用超声波、超速离心、分级盐析、分子筛层析等方法纯化HO 2。检测酶活性。结果　确定了HO 2可溶性表达最佳条件为 37℃ ,(85± 5 )r min ;确定了HO 2具体纯化路线。得到蛋白纯度为 95 .0 % ,纯化倍数为 2 .9倍 ,收得率为 4 0 .9%的活性HO 2蛋白。结论　获得了纯度高、具有活性的HO 2蛋白 ,为进一步进行HOObjective To definite the optimal expression condition and purified methods of the recombinant of heme oxygenase 2 in E.coli. Methods pMW172a/HO 2,the recombinant of heme oxygenase 2 (HO 2) cDNA with plasmid,was transformed into E.coli BL 21.Through changing the temperature and shaking speed of culture bed respectively,analyzed their influences on HO 2 expression to definite the optimal expression condition.The purification process of expressed HO 2 from E.coli BL 21 mainly included splitting bacterial 95.0%,the purification folds were 2.9 and the reclaim rate was 40.9% calculated according to the activity.Conclusion This research lay the foundation of studying the relationships between its structure and function.

关 键 词：重组血红素加氧酶-2 大肠杆菌 基因表达 纯化 

分 类 号：Q55[生物学—生物化学]