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猪外核苷酸焦磷酸酶/磷酸二酯酶1(NPP1/CD203a):猪第9工作坊簇(SWC9)单克隆抗体的NPP1/CD203a表位的克隆、转录、表达、定位及鉴定

Porcine ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1/CD203a): cloning, transcription, expression, mapping, and identification of an NPP1/CD203a epitope for swine workshop cluster 9 (SWC9) monoclonal antibodies.

作者信息

Petersen C B, Nygård A-B, Viuff B, Fredholm M, Aasted B, Salomonsen J

机构信息

Department of Veterinary Pathobiology, The Royal Veterinary and Agricultural University, DK-1870 Frederiksberg, Denmark.

出版信息

Dev Comp Immunol. 2007;31(6):618-31. doi: 10.1016/j.dci.2006.08.012. Epub 2006 Oct 9.

Abstract

Swine workshop cluster 9 (SWC9) antibody identifying a porcine epitope on macrophages and thymocytes was used to precipitate and characterize the molecule from biotinylated macrophages and to obtain peptide sequence by mass spectrometry. The protein was identified as ecto-nucleotide pyrophosphatase/phosphodiesterase 1 (NPP1/CD203a). The porcine NPP1/CD203a encoding gene was mapped to chromosome 1 using a radiation hybrid panel, and transcription was investigated by RT-PCR analysis of several tissues. The cDNA was cloned and introduced into COS7 cells resulting in expression of functionally active enzyme and verification of the specificity of an SWC9 reacting monoclonal antibody. The antibody was used for immunohistochemical examination of various porcine tissues. Most prominent expression of NPP1/CD203a was found in lung macrophages and liver sinusoids.

摘要

识别巨噬细胞和胸腺细胞上猪表位的猪车间簇9(SWC9)抗体用于从生物素化巨噬细胞中沉淀和鉴定该分子,并通过质谱获得肽序列。该蛋白质被鉴定为胞外核苷酸焦磷酸酶/磷酸二酯酶1(NPP1/CD203a)。使用辐射杂种板将猪NPP1/CD203a编码基因定位到1号染色体,并通过对多个组织的RT-PCR分析研究转录情况。克隆cDNA并将其导入COS7细胞,导致功能活性酶的表达以及SWC9反应性单克隆抗体特异性的验证。该抗体用于各种猪组织的免疫组织化学检查。在肺巨噬细胞和肝血窦中发现NPP1/CD203a的表达最为突出。

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