期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

小鼠IL-12受体β2胞内区蛋白表达及多抗制备 被引量：1

Expression of Cytoplasmic Domain of Mouse Interleukin 12 Receptor β2 and Preparation of Polyclonal Antibodies
原文传递
导出
摘要 从ConA刺激的小鼠脾细胞中抽提总RNA,用RT-PCR的方法克隆出小鼠IL-12R β2亚基胞内区基因。再将其插入到原核表达载体pGEX-4T-1中,转化宿主菌BL-21(DE3),用IPTC诱导宿主菌表达蛋白。SDS-PAGE分析结果表明,蛋白主要存在于包涵体中。利用割胶回收的方法纯化目的蛋白,免疫兔子制备多抗。经ELISA检测,证明免疫后兔血清中有高滴度的针对IL-12Rβ2亚基胞内区的抗体。 The gene of cytoplasmic domain of mouse interleukin 12 receptor (32 was amplified from ConA stimulated mouse spleen cells via RT-PCR. It was cloned into the prokaryotic expression vector pGEX-4T-1. Then the recombinant plasmid was transformed into E. coli BL-21 (DE3 ) . The fusion protein of GST-mIL-12R (32 was expressed under the condition of IPTG induction, and was found to be located in inclusion bodies. The protein was purified from SDS-PAGE gel and was used to immunize rabbit to produce polyclonal antibodies against murine IL-12R (32. When the serum antibody was measured by ELISA, it was demonstrated that the rabbit serum antibody could specifically recognize cytoplasmic domain of murine interleukin 12 receptor β2.
作者 林瑛 陈婉蓉 季永镛 孙兵
机构地区 华东师范大学生命科学学院 中国科学院上海生物化学与细胞生物学研究所
出处 《上海免疫学杂志》 CSCD 北大核心 2003年第5期293-297,共5页 Shanghai Journal of Immunology
关键词 IL-12Rβ2 基因克隆 蛋白表达 多克隆抗体 interleukin 12 receptor β2 gene cloning protein expression polyclonal antibody
分类号 R392.11 [医药卫生—免疫学]
  • 相关文献

参考文献11

  • 1J萨姆布鲁克著.分子克隆实验指南:第2版[M].北京:科学技术出版社,1992.867—897.
  • 2Trinchieri G. Interleukin-12 and the regulation of innate resistance and adaptive immunity[J]. Nat Rev Immunol, 2003,3:133.
  • 3Nagayama H, Sato K, Kawasaki H, et al. IL-12 responsiveness and expression of IL-12 receptor in human peripheral blood monocyte-derived dendritic cells[J].J Immunol,2000,165:59.
  • 4Airoldi I, Gri G, Marshall JD,et al. Expression and function of IL-12 and IL-18 receptors on human tonsillar B cells[J]. J Immunol, 2000,165 : 6880.
  • 5Chua AO, Wilkinson VL, Presky DH, et al. Cloning and characterization of a mouse IL-12 receptor-beta component [J]. J Immunoi,1995,155:4286.
  • 6Presky DH, Yang H, Minetti LJ,et al. A functional interleukin 12 receptor complex is composed of two beta-type cytokine receptor subunits[J]. Proc Natl Acad Sci USA, 1996,93:14002.
  • 7Szabo S J, Dighe AS, Gubler U, et al. Regulation of the interieukin (IL)-12R beta 2 subunit expression in developing T helper 1 (Thl)and Tn2 ceus[J].J Exp Med, 1997,185: 817.
  • 8Chakir H, Camilucci AA, Filion LG, et al. Differentiation of routine NK cells into distinct subsets based on variable expression of the IL-12R β2 subunit[J]. J Immunol, 2000,165:4985.
  • 9Wu CY, Gadina M, Wang K,et al. Cytokine regulation of IL-12 receptor beta 2 expression: differential effects on human T and NK cellsr [J].Eur J Immunol,2000,30:1364.
  • 10van Rietsehoten JG, Smits HH, van de Wetering D,et al. Silencer activity of NFATc2 in the interleukin-12 receptor beta 2 proximal promoter in human T helper cells[J]. J Biol Chem, 2001,276:34509.

共引文献11

同被引文献10

  • 1Steel DM, Whitehead AS. The major lteute phase reactants: C-reactive protein, serum amyloid P component and serum amyloidA protein. Immunol Today, 1994,15:81 - 88.
  • 2Mold C, Gresham HD, Du Clos TW. Serum amyloid Peom. ponent and C-reactive protein mediate phagocytes through routine FcyRs. Immunol,2001,166:1200 - 1205.
  • 3Pepys MB, Herbert J, Hutchinson WL, et al. Targeted pharmacological depletion of serum amyloid P component for treatment of human amyloidosis. Nature, 2002,417 : 254 - 259.
  • 4Maeda S. Use of genetically altered mice to study the role of serum amyloid P component in amyloid deposition. Amyloid, 2003,10:17 - 20.
  • 5Tashiro F, Yi S, Wakasugi S, et al. Role of serum amyloid P component for systemic amyloidosis in transgenic mice carrying human mutant transthyretin gene. Gerontology, 1991,37:56-62.
  • 6Hohenester E, Hutchinson WL, Pepys MB, et al. Crystal structure of a decameric complex of human serum amyloid P component with bound dAMP. J Mol Biol, 1997,269:570-578.
  • 7Singh PP, Kaur S. Serum amyloid P-component in murine tuberculosis:induction kinetics and intramacrophage Mycobacterium tuberculosis growth inhibition in vitro. Microbes Infect, 2006,8:541 - 551.
  • 8Ashton AW, Boehm MK, Gallimore JR, et al. Pentameric and Decameric Structures in Solution of Serum Amyloid P Component by X - ray and Neutron Scattering and Molecular Modelling Analyses. J Mol Biol, 1997,272:408 - 422.
  • 9Kim JK, Scott EA, Elbert DL, et al. Proteomic analysis of protein adsorption: Serum amyloid P adsorbs to materials and promotes leukocyte adhesion. J Biomed Mater Res A, 2005,75:199 - 209.
  • 10Pepys MB, Hawkins PN, Kahan MC, et al.Effects of Bacterial Recombinant Human C-Reactive Protein Are Caused by Contamination With Bacterial Products, Not by C-Reactive Protein Itself. Circ Res,2005,97 : e97 - 103.

引证文献1

上海免疫学杂志
2003年 第5期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部