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

人FBXO31蛋白的原核表达及抗体制备

Prokaryotic Expression of Human FBXO31 and Preparation of its Polyclonal Antibody
原文传递
导出
摘要 FBXO31位于人染色人体16q24.3,是一个乳腺癌候选抑癌基因。为了获得抗FBXO31的多克隆抗体,构建了FBXO31基因的原核表达质粒pET-28a-FBXO31。将原核表达质粒pET-28a-FBXO31转入大肠杆菌BL21(DE3)菌株中,IPTG诱导表达融合蛋白。融合蛋白主要以包涵体形式存在,常规方法纯化包涵体。将纯化的pET-28a-FBXO31融合蛋白用于免疫新西兰大白兔获得抗血清,用pET-28a-FBXO31对抗血清进行分离纯化,得到抗FBXO31多克隆抗体。用Western印迹和免疫沉淀对其进行初步鉴定。获得了兔抗人FBXO31的多克隆抗体,为进一步研究FBXO31的生物功能提供了有用的工具。 FBXO31 is the chromosome 16q24.3 senescence gene, a candidate breast tumor suppressor, and a component of an SCF complex. In order to obtain rabbit anti-FBXO31 polyclonal antibody, the complete coding sequence was cloned into pET-28a. Prokaryotic expression plasmid pET-28a-FBXO31 was constructed and expressed in the strain of E. coli BL21（DE3）. Fusion protein mainly existed in inclusion body. Fusion protein was purified and immune rabbit to obtain rabbit anti-FBXO31 polyclonal antibody. The specificity confirmed by Western blot and immunoprecipitation. The results showed that rabbit anti-FBXO31 polyclonal antibody could identify FBXO31 protein and be a useful tool in FBXO31 function analysis.
作者 黄海丽 郑文岭 赵蕊 严启涛 蔡翠霞 张宝 马文丽
机构地区 南方医科大学基因工程研究所
出处 《中国生物工程杂志》 CAS CSCD 北大核心 2010年第7期24-27,共4页 China Biotechnology
关键词 FBX031 原核表达 多克隆抗体 FBXO31 Prokaryotic expression Polyclonal antibody
分类号 Q786 [生物学—分子生物学]
  • 相关文献

参考文献10

  • 1Koegl M, Hoppe T, Schlenker S, et al. A novel ubiquitination factor, FA, is involved in muhiubiquitin chain assembly. Cell, 1999, 96(5) :635-644.
  • 2Bai C, Sen P, Hofmann K, et al. SKP1 connects cell cycle regulators to the ubiquitin proteolysis machinery through a novel motif, the F-box. Cell, 1996, 86 (2) :263-274.
  • 3Cardozo T, Pagano M. The SCF ubiquitin ligase: Insights into a molecular machine. Nat Rev Mol Cell Biol,5(9) : 739 - 753.
  • 4Yamasaki L, Pagano M. Cell cycle, proteolysis and cancer. Curr Opin Cell Biol,2004, 16(6) :623-628.
  • 5Kumar R, Neilsen P M, Crawford J, et al. FBXO31 Is the Chromosome 16q4. 3 Senescence Gene, a Candidate Breast Tumor Suppressor, and a Component of an SCF Complex. Cancer Res,2005, 65(24) : 11304-11313.
  • 6Santra M K, Wajapeyce N, Green M R. F-box protein FBXO31 mediates cyclin D1 degradation to induce G1 arrest after DNA damage. Nature, 2009, 459 (7247) :722-725.
  • 7Jia L, Sun Y. F-box proteins FBXO31 and FBX4 in regulation of eyelin D1 degradation upon DNA damage. Pigment Cell Melanoma Res, 2009,22(5 ) :518-519.
  • 8Shiloh Y. FBX031: a new player in the ever-expanding DNA damage response orchestra. Sci Signal, 2009, 2(96) : 73.
  • 9[1]萨姆布鲁克J,拉塞尔 DW.分子克隆实验指南[M].3版.北京:科学出版社,2005:1271-1374.
  • 10杨秉芬,李平,李娟,瞿秀华,李晓明,柳晓兰,孙启鸿,曹诚.兔抗人热激蛋白70样蛋白1多克隆抗体的制备与初步鉴定[J].生物技术通讯,2009,20(5):674-676. 被引量：1

二级参考文献10

  • 1Udono H,Srivastava P K.Comparison of tumor-specific immunogenicities of stress-induced proteins gp96,hsp90,and hsp70[J].J Immunology,1994,152:5398-5403.
  • 2Liu B,Ye D,Zhao Q,et al.A novel therapeutic fusion protein vaccine by two different families of heat shock proteins linked with HPV16 E7 generates potent antitumor immunity and antiangiogenesis[J].Vaccine,2008,26:1387-1396.
  • 3Zhang Y,Zan Y,Tien P,et al.Effects of heat shock protein gp96 on human dendritic cell maturation and CTL expansion[J].Biochem Biophys Res Commnn,2006,344:581-587.
  • 4Morio Y,Mayhew M,Houghton A N,et al.Induction of cellular immunity by immunization with novel hybrid peptides complexed to heat shock protein 70[J].Proc Natl Acad Sci USA,2000,97:3485-3490.
  • 5Flechtne J B,Cohane K P,Andjelic S,et al.High-affinity interactions between peptides and heat shock protein 70 augment CD8^+ T lymphocyte immune responses[J].J Immunol,2006,177:1017-1027.
  • 6Wan Tan,Zhou Xiangyang,Cao Xuetao,et al.Novel heat shock protein Hsp70L1 activates dendritic cells and acts as a Th1 polarizing adjuvant[J].Blood,2004,103:1747-1754.
  • 7Wu Y,Wan T,Cao X,et al.Hsp70-like protein 1 fusion protein enhances induction of carcinoembryonic antigen-specific CD8^+ CTL response by dendritic cell vaccine[J].Cancer Res,2005,65:4947-4954.
  • 8Otto H,Conz C,Rospert S,et al.The chaperones MPP11 and HSP70L1 form the mammalian ribosome-associated complex[J].Proc Natl Acad Sci USA,2005,102:10064-10069.
  • 9Kim Hyun-Eui,Jiang Xnejun,Wang Xiaodong,et al.PHAPI,CAS,and Hsp70 promote apoptosome formation by preventing Apaf-1 aggregation and enhancing nucleotide exchange on Apaf-1[J].Mol Cell,2008,30:239-247.
  • 10Gurbuxani S,Schmitt E,Garrido C,et al.Heat shock protein 70 binding inhibits the nuclear import of apoptosis-indueing factor[J].Oncogene,2003,22:6669-6678.

共引文献1

中国生物工程杂志
2010年 第7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