| Gene cloning of human soluble CD14 and its expression ineucaryotic cells |
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| 引用本文： | 荫俊,白洁,王威,宋伟,王忠泽. Gene cloning of human soluble CD14 and its expression ineucaryotic cells[J]. 中华创伤杂志(英文版), 2002, 5(3): 156-160 |
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| 作者姓名： | 荫俊 白洁 王威 宋伟 王忠泽 |
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| 作者单位： | BeijingInstituteofMicrobiology,20,Dongdajie,Fengtai,Beijing100071,China |
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| 摘 要： | Objective: To express human soluble CD14 (sCD14) in eukaryotic cens. Methods : Human sCD14 cDNA was amplified from U937 cells with RT-PCR method. The recombinant expression plasmid pEFI/HisC/sCD14^348ss was constructed and the expression in COS-7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno-affinity chromatography. Results: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical, sCD14 was expressed with highyield. The expressed product was purified to above 90%.Recombinant sCDI4, specifically combinable with endotoxins, had a natural biological activity. Conclusions: Human sCD14 was expressed in COS-7 cells, which laid a foundation for further study.
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| 关 键 词： | 基因克隆 可溶性CD14 基因表达 序列分析 COS-7细胞 |
Gene cloning of human soluble CD14 and its expression in eucaryotic cells |
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| Jun Yin,Jie Bai,Wei Wang,Wei Song,Zhongze Wang. Gene cloning of human soluble CD14 and its expression in eucaryotic cells[J]. Chinese journal of traumatology, 2002, 5(3): 156-160 |
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| Authors: | Jun Yin Jie Bai Wei Wang Wei Song Zhongze Wang |
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| Affiliation: | Beijing Institute of Microbiology, 20, Dongdajie, Fengtai, Beijing 100071, China |
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| Abstract: | OBJECTIVE: To express human soluble CD14 (sCD14) in eukaryotic cells. METHODS: Human sCD14 cDNA was amplified from U937 cells with RT-PCR method. The recombinant expression plasmid pEF1/HisC/sCD14(348aa) was constructed and the expression in COS-7 cells was carried out using liposome transfection method. The yield was examined with scanning map identification. The expressed product was purified by immuno-affinity chromatography. RESULTS: Sequence analysis demonstrated that the amplified gene sequence and those reported by documents were completely identical. sCD14 was expressed with high-yield. The expressed product was purified to above 90%. Recombinant sCD14, specifically combinable with endotoxins, had a natural biological activity. CONCLUSIONS: Human sCD14 was expressed in COS-7 cells, which laid a foundation for further study. |
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| Keywords: | CD Gene Eucaryotic expression Biological function LPS |
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