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[白细胞介素-1受体拮抗剂-Fcε融合基因的克隆、表达及鉴定]

[Cloning, expression and identification of IL-1ra-Fcepsilon fusion gene].

作者信息

Liu Zhongcheng, Zou Minji, Wang Yuanyuan, Wang Jiaxi, Xu Donggang

机构信息

College of Pharmacy, Hebei University, Baoding 071002, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2008 Oct;24(10):1754-60.

Abstract

Both interleukin-1 and IgE are important in the pathogenic mechanism of the allergy asthma. cDNA of interleukin-1receptor antagonist (IL-1ra) and IgE were cloned and a prokaryotic expression vector IL-1ra-Fcepsilon/pBV220 was constructed. The vector was transformed into Escherichia coli BL21(DE3). The fusion protein was expressed successfully in the form of inclusion body. The recombination protein of IL-1ra-Fcepsilon was highly purified by chromatography of gel filtration and ion exchange, which was identifited by Western blotting. The cell assay showed that the activity of IL-1ra-Fcepsilon was as high as IL-1ra in vitro after refolding. The pharmacokenetic profile of IL-1ra-Fcepsilon and L-1ra was analyzed, and the half time of IL-1ra-Fcepsilon is 4.78 times than that of IL-1ra.

摘要

白细胞介素-1和免疫球蛋白E在过敏性哮喘的发病机制中都很重要。克隆了白细胞介素-1受体拮抗剂(IL-1ra)和免疫球蛋白E的cDNA,并构建了原核表达载体IL-1ra-Fcepsilon/pBV220。将该载体转化到大肠杆菌BL21(DE3)中。融合蛋白以包涵体的形式成功表达。通过凝胶过滤和离子交换层析对IL-1ra-Fcepsilon重组蛋白进行了高度纯化,并通过蛋白质印迹法进行鉴定。细胞试验表明,复性后的IL-1ra-Fcepsilon在体外的活性与IL-1ra一样高。分析了IL-1ra-Fcepsilon和L-1ra的药代动力学特征,IL-1ra-Fcepsilon的半衰期是IL-1ra的4.78倍。

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