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条斑紫菜锰超氧化物歧化酶原核表达及多克隆抗体制备
引用本文:邵林,茅云翔,阎斌伦,孔凡娜,王健.条斑紫菜锰超氧化物歧化酶原核表达及多克隆抗体制备[J].中国海洋大学学报(自然科学版),2010,40(4).
作者姓名:邵林  茅云翔  阎斌伦  孔凡娜  王健
作者单位:1. 中国海洋大学海洋生命学院,山东,青岛,266003
2. 淮海工学院江苏省海洋生物技术重点建设实验室,江苏,连云港,222005
基金项目:国家高技术研究发展计划项目,国家自然科学基金项目(30972247)教育部新世纪人才支持计划,教育部科学技术研究重点项目,江苏省海洋生物技术重点建设实验室开放课题资助 
摘    要:研究紫菜抗逆的分子机制,将本实验室克隆的条斑紫菜(Porphyra yezoensisUeda)锰超氧化物歧化酶(manganese superoxide dismutase,Mn SOD)基因全长cDNA克隆到质粒pET-30c(+)中,构建原核表达载体pET-S,转化大肠杆菌BL21(DE3)表达Mn SOD,表达产物的分子量约为30 KDa。利用不同浓度的Mn2+诱导重组Mn SOD的表达,可以检测到比活力随Mn2+浓度增高而上升的趋势。用重组Mn SOD免疫新西兰大白兔,制备了多克隆抗体,多克隆抗体的效价为1∶8 000。免疫印迹实验(Western Blot)表明该多克隆抗体具有很好的特异性。

关 键 词:条斑紫菜  锰超氧化物歧化酶(Mn  原核表达  酶活性  多克隆抗体

Expression and Polyclonal Antibody Preparation of Prokaryotic Recombinant Manganese Superoxide Dismutase (Mn SOD) fromPorphyra yezoensisUeda
SHAO Lin,MAO Yun-Xiang,YAN Bin-Lun,KONG Fan-Na,WANG Jian.Expression and Polyclonal Antibody Preparation of Prokaryotic Recombinant Manganese Superoxide Dismutase (Mn SOD) fromPorphyra yezoensisUeda[J].Periodical of Ocean University of China,2010,40(4).
Authors:SHAO Lin  MAO Yun-Xiang  YAN Bin-Lun  KONG Fan-Na  WANG Jian
Institution:SHAO+Lin1,MAO+Yun-Xiang1,YAN+Bin-Lun2,KONG+Fan-Na1,WANG+Jian1(1.College+of+Marine+Life+Science,Ocean+University+of+China,Qingdao+266003,China,2.Jiangsu+Key+Laboratory+of+Marine+Biotechnology,Huaihai+Institute+of+Technology,Lianyungang+222005,China)
Abstract:A cDNA corresponding to manganese superoxide dismutase(SOD; EC 1.15.1.1.) from Porphyra yezoensis Ueda was overexpressed in the Escherichia coli BL21(DE3) using the expression vector pET-30c(+) and the molecular weight of the recombinant MnSOD is about 30 KDa. With the concentration of Mn~(2+) increasing, the activity of the recombinant MnSOD is enhanced by inducement of Mn~(2+). The recombinant Mn SOD was used to generate a polyclonal antibody in New Zealand white rabbits that recognized Mn SOD and the value is 1∶8 000. The western blot shows the polyclonal antibody can be used effectively.
Keywords:SOD)  Porphyra yezoensis Ueda  manganese superoxide dismutase(Mn SOD)  prokaryotic expression  enzymatic activity  polyclonal antibody
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