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人FcγRIIb胞外区原核表达载体的构建及表达

Keywords: sFcγRIIb基因,载体构建,诱导表达,纯化

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Abstract:

目的构建人FcγRIIb胞外区蛋白原核表达载体pET-sFcγRIIb,诱导表达并纯化。方法PCR扩增获得人sFcγRIIb基因,并将其定向克隆至原核表达载体pET-32a(+),构建原核表达重组质粒pET-sFcγRIIb,经PCR、限制性内切酶鉴定及测序分析后,转化大肠杆菌BL21,采用不同温度和时间的IPTG诱导蛋白表达,确定最佳诱导条件,SDS―PAGE分析正确后,用镍琼脂糖磁珠纯化。结果扩增出了人sFcγRIIb基因,成功构建原核表达重组质粒pET-sFcγRIIb,30℃10hIPTG诱导表达并纯化出41.5kDa的融合蛋白。结论获得重组人FcγRIIb胞外区蛋白,为以后人FcγRIIb胞外区蛋白功能的深入研究奠定基础。

References

[1]  Tatyana T,Jonathan AD,Silvia B. FcgRIIB as a modulator of autoimmune disease susceptibility [J]. Autoimmunity,2007,40(6):409-410.
[2]  Sautes C, Galinha A,Bouchard C,et al.Recombinant soluble Fc gamma receptors: production, purification and biological activities[J].Chromatogr B Biomed Appl,1994,662:197-207.
[3]  Huizinga TW,van der Schoot CE,Jost C,et al. The PI-linked receptor FcRIII is released on stimulation of neutrophils [J]. Nature,1988,333:667-669.
[4]  Gavin AL,Wines BD,Powell MS,et al.Recombinant soluble Fc gamma RII inhibits immune complex precipitation[J].Clin Exp Immunol,1995,102:620-625.
[5]  Ierino FL,Powell MS, McKenzie IF,et al. Recombinant soluble human Fcgamma RII: production, characterization, and inhibition of the Arthus reaction [J]. Exp Med,1993,178:1617?1628.
[6]  Werwitzke S,Trick D,Sondermann P,et al.Treatment of Lupus-prone NZB/NZW F1 Mice with Recombinant Soluble FcγReceptor II(CD32)[J].Ann Rheum Dis,2008,67(2):154-161.
[7]  Nimmerjahn F,Ravetch JV. Fc gamma receptors: old friends and new family members[J]. Immunity, 2006,24(1):19-28.
[8]  Horejs HJ,Hren A,Mudde GC,et al. Inhibition of immunoglobulin E synthesis through Fc gammaRII(CD32)by a mechanism independent of B-cell receptor co-cross-linking [J].Immunology,2005,115(3): 407-415.

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