| 牛冠状病毒4H12/1D4单链基因片段在大肠杆菌中的表达 |
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| 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, 0(1): 38-40 |
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| 作者姓名： | 李学伍 陈焕春 |
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| 作者单位： | [1]河南省农业科学院 [2]华中农业大学 |
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| 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％
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| 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 |
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| 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, 0(1): 38-40 |
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| Authors: | Li Xuewu Yang Yanyan |
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| 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. |
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| Keywords: | Bovine coronavirus Genetically engineered antibody |
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