| HLA-B*2705重链的原核表达和活性鉴定 |
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| 引用本文： | 李彦博,孙玉英,郭斯启,孙业平,张秀云,孔繁华,奚永志.HLA-B*2705重链的原核表达和活性鉴定[J].中国实验血液学杂志,2007,15(5):1028-1031. |
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| 作者姓名： | 李彦博 孙玉英 郭斯启 孙业平 张秀云 孔繁华 奚永志 |
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| 作者单位： | 军事医学科学院附属医院免疫学研究室国家生物医学分析中心免疫分析实验室,北京,100071 |
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| 基金项目： | 国家自然科学基金;军队医药卫生科研项目 |
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| 摘 要： | 本研究探讨HLA-B*2705重链的原核表达及活性鉴定。从HLA-B*2705全长cDNA中PCR扩增出膜外区片段并克隆进入pGEM-T载体,序列测定后构建原核融合表达载体pET32a( )-B*2705,并转化大肠杆菌。Western Blot和阻断抗体反应鉴定它们在体外是否具有HLA-B27抗原活性。结果表明:HLA-B*2705重链融合蛋白在大肠杆菌中获得高效融合表达,表达产量占细菌总蛋白的50%以上;通过Western Blot和阻断抗体反应鉴定它们在体外具有HLA-B27抗原活性。结论:本研究获得了HLA-B*2705重链融合蛋白,为今后的相关研究打下了基础。
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| 关 键 词： | HLA-B*2705重链 原核表达 抗原活性 |
| 文章编号： | 1009-2137（2007）05-1028-04 |
| 修稿时间： | 2006-11-01 |
Prokaryotic Expression of HLA-B*2705 Heavy Chain and Identification for Its Activity |
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| LI Yan-Bo,SUN Yu-Ying,GUO Si-Qi,SUN Ye-Ping,ZHANG Xiu-Yun,KONG Fan-Hua,XI Yong-Zhi.Prokaryotic Expression of HLA-B*2705 Heavy Chain and Identification for Its Activity[J].Journal of Experimental Hematology,2007,15(5):1028-1031. |
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| Authors: | LI Yan-Bo SUN Yu-Ying GUO Si-Qi SUN Ye-Ping ZHANG Xiu-Yun KONG Fan-Hua XI Yong-Zhi |
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| Institution: | Department of Immunology, Affiliated Hospital of Academy of Military Medical Sciences, Laboratory of lmmunoassy, National Center of Biomedical Analysis, Beijing 100039, China |
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| Abstract: | In order to investigate the expression of heavy chain of HLA-B*2705 in prokaryotic system and identify its activity, the extra-membrane gene fragment of HLA-B*2705 was amplified from full-length HLA-B*2705 cDNA by PCR and cloned into pGEM-T vector. After identification by sequencing, the prokaryotic expressing vector pET32a( )-B*2705 was constructed. The antigenic activity of expressed protein was identified by Western blot and antibody blocking reaction. The results indicated that the fused HLA-B*2705 protein expression with high efficiency was obtained. The expressed product was more than 50% of the total bacteria protein. The antigenic activity of expressed protein was confirmed by Western blot and antibody blocking reaction. It is concluded that HLA-B*2705 fusion protein are obtained as basis for the further studies. |
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| Keywords: | HLA-B*2705 heavy chain prokaryotic expression antigenic activity |
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