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

类朊蛋白Shadoo基因在Neuro-2α中的表达及细胞定位分析

Shadoo gene expression level and location in Neuro-2a
原文传递
导出
摘要 为构建鼠类朊蛋白Shadoo基因重组真核表达质粒,并分析类朊蛋白Shadoo在鼠神经瘤细胞(Neuro-2α)内表达及定位。采用PCR方法把Flag标签序列插入到Shadoo阅读框122与123氨基酸位点之间,构建Shadoo-Flag融合基因片段,把Shadoo-Flag片段插入到真核表达质粒pcDNA3.1,构建重组真核表达pcDNA3.1-Shadoo-Flag,质粒酶切、测序鉴定正确后,脂质体法转入Neuro-2α细胞,检测Shadoo基因表达及表达部位。结果重组真核表达质粒pcDNA3.1-Shadoo-Flag转入Neuro-2α细胞,能够正确表达出相对分子质量为12 000的Shadoo蛋白,并定位于细胞膜。 To construct the recombinant eukaryotic expression plasmid pcDNA3. 1-Shadoo-Flag, Shadoo ORF with Flag epitope tag （8 aa） inserted between 122-123 of ORF by PCR method, pcD-NA3.1-Shadoo-Flag was cloned with Shadoo-Flag through HindⅢ and BamH Ⅰ . pcDNA3.1-Sha-doo-Flag was verified by double enzyme digestion and sequencing. Shadoo expression level and lo- cation in Neuro-2a were investigated after the constructed eukaryotic expression plamid was trans- fected to Neuro-2a cell line by lipofectamin method. The constructed eukaryotic expression plamid was transfected to Neuro-2α cell line by lipofectamin method,and fused protein in eukaryotic cell was detected by immunocytochemical staining as well as Western blot. Recombinant eukaryotic ex-pression plasmid pcDNA3. 1-Shadoo-Flag was successfully contructed. Shadoo fusion protein was expressed in Neuro-2α cell line and Shadoo is located on the membrane of Neuro-2α cell. It estab-lishs the foundation for future research on Shadoo gene function in TSE.
作者 曹德康 王雅静 万家余 李忠义 孟轲音 钱军
机构地区 武装警察部队后勤部疾病预防控制中心 武警总医院 军事医学科学院军事兽医研究所吉林省人兽共患病预防与控制重点实验室
出处 《中国兽医学报》 CAS CSCD 北大核心 2013年第8期1223-1226,共4页 Chinese Journal of Veterinary Science
基金 国家自然科学基金资助项目(30972197 31072148 31101782) 吉林省科技发展计划资助项目(201105038 201215096)
关键词 朊蛋白 Shadoo基因 鼠神经癌细胞株 PrP^c Shadoo gene Neuro-2
分类号 S855.3 [农业科学—临床兽医学] R535 [医药卫生—内科学]
  • 相关文献

参考文献12

  • 1Kaneko K, Zulianello L, Scott M, et al. Evidence for protein X binding to a discontinuous epitope on the cellular prion protein during scrapie prion propagation.
  • 2Proc Natl Acad Sci USA, 1997,94: 10069-10074.
  • 3Aguzzi A,Baumann Fv BrernerJ. The prion's elusive reason for being[J]. Annu Rev Neurosci , 2008,31: 439-477.
  • 4Edenhofer F, Rieger R, Famulok M, et al. Prion pro?tein PrP' interacts with molecular chaperones of the Hsp60 family[J].J Virol, 1996,70: 4724-4728.
  • 5Hundt Cv PeyrinJ Mv Haik Sv et al. Identification of interaction domains of the prion protein with its 37- kDa/67-kDa laminin receptor[J]. EMBOJ ,2001,20: 5876-5886.
  • 6Rieger R, Edenhofer F, Lasmezas C I, et al. The hu?man 37-kDa laminin receptor precursor interacts with the prion protein in eukaryotic cells[J]. Nat Med, 1997(3):1383-1388.
  • 7Maness P F, Schachner M. Neural recognition mole?cules of the immunoglobulin superfamily: signaling transducers of axon guidance and neuronal migration[J]. Nat Neuro Sci,2007, 10: 19-26.
  • 8Schmitt-Ulms G, Legname G, Baldwin M A, et al. Binding of neural cell adhesion molecules (N-CAMs) to the cellular prion protein[J].J Mol Bioi, 2001 , 314: 1209-1925.
  • 9Premzl M, Sangiorgio L, Strumbo B. Shadoo , a new protein highly conserved from fish to mammals and with similarity to prion protein[J]. Gene, 2003,314: 89-102.
  • 10Uboldi C, Paulis M, Guidi E. Cloning of the bovine prion-like Shadoo (SPRN) gene by comparative anal?ysis of the predicted genomic locus[J]. Mamm Ge?nome, 2006,17: 1130-1139.
中国兽医学报
2013年 第8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