| 人CD20跨膜区与g3pN端融合基因的克隆及其在大肠杆菌中的表达 |
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| 引用本文： | 张效云,孙志伟,俞炜源,程建贞.人CD20跨膜区与g3pN端融合基因的克隆及其在大肠杆菌中的表达[J].细胞与分子免疫学杂志,2004,20(4):481-483. |
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| 作者姓名： | 张效云 孙志伟 俞炜源 程建贞 |
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| 作者单位： | 1. 河北北方学院生物化学教研室,河北,张家口,075000
2. 军事医学科学院生物工程研究所,北京,100071 |
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| 摘 要： | 目的 :构建g3pN1 hCD2 0跨膜及胞外区基因表达载体 ,并在大肠杆菌中进行高效融合表达。方法 :利用反转录PCR和PCR方法 ,分别从Daudi细胞和M13K0 7噬菌体中扩增hCD2 0基因和编码g3pN1蛋白N1端的基因 ,并重组到pTIG Trx表达载体中 ,在大肠杆菌中融合表达。结果 :表达产物可被抗CD2 0分子的单克隆抗体 (mAb)识别。结论 :成功地表达并鉴定了目的蛋白。
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| 关 键 词： | CD20基因 Daudi细胞 g3p 噬菌体抗体库 |
| 文章编号： | 1007-8738(2004)04-0481-03 |
| 修稿时间： | 2003年10月20 |
Cloning and expression of fusion gene of transmembrane domain of human CD20 and g3pN in Escherichia coli |
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| ZHANG Xiao-yun,SUN Zhi-wei,YU Wei-yuan,CHENG Jian-zhen.Cloning and expression of fusion gene of transmembrane domain of human CD20 and g3pN in Escherichia coli[J].Journal of Cellular and Molecular Immunology,2004,20(4):481-483. |
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| Authors: | ZHANG Xiao-yun SUN Zhi-wei YU Wei-yuan CHENG Jian-zhen |
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| Affiliation: | Department of Biochemistry, Hebei North College, Zhangjiakou 075000, China. xyz700928@yahoo.com.cn |
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| Abstract: | AIM: To construct the expression vector containing transmembrane domain gene of human CD20 and g3pN1 gene and express the fusion gene high-efficiently in E.coli. METHODS: The human CD20 gene and g3pN1 domain gene were amplified by RT-PCR and PCR from Daudi cells and M13K07 phage antibody library, respectively, and then cloned into expression vector pTIG-Trx.The constructed expression vector was expressed in E.coli. RESULTS: Western blot analysis showed that expressed product could bind to anti-CD20 mAb. CONCLUSION: The pTIG-GS has been constructed and expressed successfully in E.coli, which lays the foundation for further screening anti-CD20 antibody from phage antibody library. |
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| Keywords: | CD20 gene Daudi cell g3p phage antibody library |
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