| Mn-SOD与抗癌胚抗原单链抗体基因的融合及高效表达 |
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| 引用本文： | 贺华君,杨卫东,常雅宁,施惠娟,杨冠珍,吴祥甫,袁勤生.Mn-SOD与抗癌胚抗原单链抗体基因的融合及高效表达[J].生物工程学报,2000,16(5):566-569. |
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| 作者姓名： | 贺华君 杨卫东 常雅宁 施惠娟 杨冠珍 吴祥甫 袁勤生 |
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| 作者单位： | [1]华东理工大学生物化学研究所,上海 [2]中国科学院上海生物化学研究所,上海 |
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| 摘 要： | 将Ｍｎ－ＳＯＤ与抗癌胚抗原（ＣＥＡ）单链抗体基因（Ｓｃ－Ｆｖ ｇｅｎｅ）融合，重组到含Ｔ７启动子的表达载体ｐＥＴ－２２ｂ（＋）中，构建表达质粒ｐＥＴＭｎ－ＳＯＤ－ＳｃＦｖ，并转化大肠杆菌ＢＬ２１（ＤＥ３），进行高效表达，表达物占菌体可溶性总蛋白的２４％。ＳＤＳ－ＰＡＧＥ和蛋白质和迹图谱显示表达物分子量为４５ｋＤ与融合基因编码蛋白质的理论值相符。该蛋白质在大肠杆菌中为泌型表达有利于纯化。ＲＩＡ测定表
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| 关 键 词： | 癌胚抗原 单链抗体 Mn－SOD 融合表达 肿瘤治疗 |
| 文章编号： | 1000-3061(2000)05-0566-04 |
| 修稿时间： | 1999-12-05 |
Fusion and Expression of the Gene Encoding Human Mn-SOD to Anti-CEA Single-chain antibody in Escherichia coli |
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| HE Hua-Jun
YANG Wei-Dong
CHANG Ya-Ning
,SHI Hui-Juan
YANG Guan-Zhen
,WU Xiang-Fu.Fusion and Expression of the Gene Encoding Human Mn-SOD to Anti-CEA Single-chain antibody in Escherichia coli[J].Chinese Journal of Biotechnology,2000,16(5):566-569. |
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| Authors: | HE Hua-Jun
YANG Wei-Dong
CHANG Ya-Ning SHI Hui-Juan
YANG Guan-Zhen WU Xiang-Fu |
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| Institution: | HE Hua-Jun
;(Institute of Biochemistry, East China University of Science and Technology, Shanghai 200237 Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031);YANG Wei-Dong
;(Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031);CHANG Ya-Ning
,SHI Hui-Juan
;(Institute of Biochemistry, East China University of Science and Technology, Shanghai 200237);YANG Guan-Zhen
,WU Xiang-Fu
;(Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031) |
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| Abstract: | The gene encoding human manganese\|superoxide dismutase (Mn\|SOD) was fused to anti\|carcinoembryonic antigen single\|chain antibody gene to construct the fusion gene,then was ligated into prokaryotic expression vector pET\|22b( ),The fusion gene was expressed in E.coli at high level,accounting for 24% of the total bacteria soluble protein;and was characterized by SDS\|PAGE and Western\|blot analysis;the expression product had the CEA\|binding ability in RIA,and also had the SOD activity by pyrogallol autoxidation assay.So,the Mn\|SOD moiety retains substantial enzymatic activity,where the ScFv moiety can deliver the fusion protein to tumor,Mn\|SOD is a potential tumor\|suppressor gene,maybe the fusion protein can provide a new pathway to tumor therapy. |
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| Keywords: | Carcinoembryonic antigen single-chain antibody manganese-superoxide dismutase fusion expression |
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