| 人可溶性CD14基因克隆表达及功能研究 |
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| 引用本文： | 白洁,荫俊,王占东,王威,王忠泽,宋伟.人可溶性CD14基因克隆表达及功能研究[J].生物工程学报,2001,17(3):269-272. |
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| 作者姓名： | 白洁 荫俊 王占东 王威 王忠泽 宋伟 |
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| 作者单位： | 1. 白求恩医科大学第二临床医院
2. 北京微生物流行病研究所 |
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| 基金项目： | 国家自然科学基金资助项目(39870316). |
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| 摘 要： | 利用反转录PCR技术，从U937细胞总RNA中，扩增编码人可溶性CD14的基因序列，构建了重组表达质粒pEF1/HisC/sCD14^348aa；用脂质体转染法，实现了在真核细胞中的高效表达；用免疫亲和层析纯化表达产物，纯度达90％以上；PLS刺激U937细胞产生CD14的变化，证明了表达产物具有结合LPS的功能。
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| 关 键 词： | CD14 基因克隆 序列分析 真核表达 纯化 生物学功能 |
| 文章编号： | 1000-3061(2001)03-0269-04 |
| 修稿时间： | 2000年9月25日 |
Clone and expression of human soluble CD14 and study of its function] |
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| J Bai,J Yin,Z D Wang,W Wang,Z Z Wang,W Song.Clone and expression of human soluble CD14 and study of its function][J].Chinese Journal of Biotechnology,2001,17(3):269-272. |
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| Authors: | J Bai J Yin Z D Wang W Wang Z Z Wang W Song |
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| Institution: | Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China. haku@263.net |
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| Abstract: | Human soluble CD14(sCD14) cDNA fragment was amplified using total RNA extracted from U937 cells by RT-PCR of sCD14 gene, and the recombinant expression plasmid pEF1/HisC/sCD14 348aa was constructed. Then the expression in eukaryotic cell was carry out by liposome transfection method. It demonstrated that the expression level was relatively high by scanning map identification. The expressed product was purified by immunoaffinity chromatography and the purity was above 90%. The changes of CD14 brought by LPS stimulating U937 cell proved the product had the function of combine with LPS. |
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