| 顶头孢霉cefG基因的表达及抗体制备的研究 |
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| 引用本文： | 程作,李强,王文雅,袁其朋. 顶头孢霉cefG基因的表达及抗体制备的研究[J]. 北京化工大学学报(自然科学版), 2010, 37(5): 107-110. DOI: 10.3969/j.issn.1671-4628.2010.05.022 |
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| 作者姓名： | 程作 李强 王文雅 袁其朋 |
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| 作者单位： | 1. 北京化工大学 生命科学与技术学院， 北京 100029; 2. 清华大学 化学工程系， 北京100084 |
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| 基金项目： | 国家"863"计划 |
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| 摘 要： | 通过PCR方法得到带酶切位点的cefG基因,用于构建表达质粒pET30-G,并在大肠杆菌里成功表达出乙酰转移酶的包涵体。将该重组蛋白的标准品作为抗原,免疫BALB/c小鼠得到多克隆抗体,酶联免疫吸附测定(ELISA)分析得抗体的效价满足实验需求。利用该抗体绘制出抗原浓度的标准曲线,确定待测抗原适合的检测浓度在0.16μg/mL以下。制得的抗体在重组大肠杆菌里检测到了乙酰转移酶,证明所得抗体具有实验价值。
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| 关 键 词： | 顶头孢霉 cefG 多克隆抗体 酶联免疫吸附测定 |
| 收稿时间： | 2010-04-14 |
Recombinant expression and antibody preparation of cefG by Acremonium chrysogenum |
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| CHENG Zuo,LI Qiang,WANG WenYa,YUAN QiPeng. Recombinant expression and antibody preparation of cefG by Acremonium chrysogenum[J]. Journal of Beijing University of Chemical Technology, 2010, 37(5): 107-110. DOI: 10.3969/j.issn.1671-4628.2010.05.022 |
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| Authors: | CHENG Zuo LI Qiang WANG WenYa YUAN QiPeng |
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| Affiliation: | 1. College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029; 2. Department of Chemical Engineering, Tsinghua University, Beijing 100084, China |
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| Abstract: | Amplification by the polymerase chain reaction(PCR) of the cefG gene with a restriction enzyme cutting site has been used to construct the expression plasmid pET30-G,which expressed acetyltransferase in E.coli in the form of inclusion bodies.The recombinant protein was then vaccinated into BALB/c mice to obtain polyclonal antibodies.Satisfactory antibody titers were obtained by ELISA analysis.The calibration curve of the antigen concentration was drawn using this antibody,and indicated that the optimum conc... |
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| Keywords: | cefG |
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