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| BP180NC16A-IgG1Fc融合蛋白真核表达体系构建 | |
| 引用本文： | 王佩茹,王明悦,朱学骏,陈喜雪. BP180NC16A-IgG1Fc融合蛋白真核表达体系构建[J]. 中国皮肤性病学杂志, 2012, 26(9): 797-800 |
| 作者姓名： | 王佩茹 王明悦 朱学骏 陈喜雪 |
| 作者单位： | 北京大学第一医院皮肤性病科,北京,100034 |
| 基金项目： | 高等学校博士学科点专项科研基金新教师基金课题，国家自然科学基金青年科学基金项目 |
| 摘 要： | 目的构建BP180NC16A-IgG1Fc融合蛋白的真核表达体系。方法①以逆转录PCR(reverse transcrip-tion-polymerase chain reaction,RT-PCR)方法从表皮组织提取总RNA中扩增编码人BP180NC16A的cDNA,与包含编码人免疫球蛋白IgG1Fc段恒定区基因的质粒pFUSE-hIgG1e4-Fc2融合,形成重组pFUSE-hIgG1e4-Fc2-BP180NC16A质粒。②重组质粒转染真核HEK293T细胞,48h后收集细胞培养上清。③通过免疫印迹的方法鉴定转染细胞上清中分泌的BP180NC16A-IgG1Fc融合蛋白。结果构建了pFUSE-hIgG1e4-Fc2-BP180NC16A真核表达质粒并在HEK293T细胞成功表达BP180NC16A-IgG1Fc融合蛋白。结论 BP180NC16A-IgG1Fc融合蛋白能够成功真核表达。 |
| 关 键 词： | BP180NC16A IgG1Fc 融合蛋白 真核表达 |
Eukaryotic Expression of BP180NC16A-IgG1Fc Fusion Protein |
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| WANG Pei-ru , WANG Ming-yue , ZHU Xue-jun , CHEN Xi-xue. Eukaryotic Expression of BP180NC16A-IgG1Fc Fusion Protein[J]. The Chinese Journal of Dermatovenereology, 2012, 26(9): 797-800 | |
| Authors: | WANG Pei-ru WANG Ming-yue ZHU Xue-jun CHEN Xi-xue |
| Affiliation: | (Department of Dermatology and Venereology,Peking University First Hospital,Beijing 100034,China) |
| Abstract: | Objective To construct eukaryotic expression system of BP180NC16A-IgG1Fc fusion protein.Methods We cloned BP180NC16A cDNA from total RNA extracted from the human epidermis by RT-PCR, then linked BP180NC16A cDNA with plasmid pFUSE-hIgG1e4-Fc2 which encodes human IgG1Fc constant region, to construct recombinant plasmid pFUSE-hIgG1e4-Fc2-BP180NC16A. Recombinant plasmids were then transfected in HEK 293T cells. Finally, we detected the BP180NC16A-IgG1Fc fusion protein in the supernatant by western blotting. Results HEK293T cells successfully expressed BP180NC16A-IgG1Fc fusion protein. Conclusion BP180NC16A-IgG1Fc fusion protein can be eukaryotically expressed. |
| Keywords: | BP180NC16A IgG1Fc Fusion protein Eukaryotic expression |
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