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牛冠状病毒4H12/1D4单链基因片段在大肠杆菌中的表达
引用本文:李学伍,陈焕春.牛冠状病毒4H12/1D4单链基因片段在大肠杆菌中的表达[J].河南农业科学,1997(1):38-40.
作者姓名:李学伍  陈焕春
作者单位:[1]河南省农业科学院 [2]华中农业大学
摘    要:用EcoRI酶切PR5B4质粒,采用“V”字形内槽电洗脱法回收约0.804Kb牛冠状病毒基因工程抗体4H12/1D4单链基因片段,并将其插入载体质粒PET-17b的EcoRI位点构成重组质粒PET-D4,将重组质粒转化DH5α感受态细胞,在IPTG的诱导下表达,细菌裂解物经SDS-PAGE电泳筛选,发现含PET-D4细菌经诱导新增一条约28KDα蛋白带,该蛋白带与设计基因工程抗体的大小相符,经光密度扫描,表达量约占菌体总蛋白的14%

关 键 词:牛冠状病毒  基因工程抗体

Expression of a Gene Fragment of the Monoclonal Antibody 4H12/1D4 to Bovine Coronavirus in Escherichia Coli
Li Xuewu,Yang Yanyan.Expression of a Gene Fragment of the Monoclonal Antibody 4H12/1D4 to Bovine Coronavirus in Escherichia Coli[J].Journal of Henan Agricultural Sciences,1997(1):38-40.
Authors:Li Xuewu  Yang Yanyan
Abstract:The recombinant plasmid PR5b4 was digested with EcoRI and an insert of 0.804 kb containing a single chain gene fragment of the monoclonal antibody 4H12/1D4 to bovine coronavirus was recovered by electrophoresis.The fragment was inserted in plasmid PET-17 b pre-digested with EcoRI and the recombinant was named as PET-D4.In the presence of the inducer IPTG the E.coli-DH5α transformed with PET-D4 expressed a peptide with apparent molecular weight of 28 kDa on SDS-PAGE,being consistent with that designed.The 28 kDa peptide took 14% of the total cellular protein in the transformed E.coli-DH5α.
Keywords:Bovine coronavirus\ Genetically engineered antibody
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