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Soluble expression of NDRG2 protein in pichia yeast and purification of the expression product
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- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 12
- DOI:
- 10.3321/j.issn:1000-2790.2003.12.002
- Key Word:
- NDRG2蛋白;毕赤酵母;基因表达;亲合纯化
Abstract： 目的:酵母表达并纯化可溶性NDRG2蛋白.方法: 从pRSET-A-ndrg2重组质粒中扩增出6his-ndrg2融合基因,克隆入酵母表达载体pPIC3.5K;酵母重组表达载体pPIC3.5K-6his-ndrg2电转化入毕赤酵母GS115,并进行营养缺陷筛选和G418抗性筛选;对高拷贝整合的重组酵母表达菌株诱导表达,可溶性表达产物经Ni-NTA亲合层析纯化.结果:构建得到酵母融合重组表达载体pPIC3.5K-6his-ndrg2;经G418浓度梯度筛选得到串联整合16个重组表达载体以上的重组酵母表达菌株;诱导后表达得到6his-NDRG2融合蛋白,并成功纯化得到可溶性表达产物.结论: 表达并纯化得到可溶性NDRG2蛋白,为进一步研究NDRG2的结构与功能打下了必要的基础.
- [1]杨敏,刘新平,韩炯,等.乙型肝炎病毒PreS基因的克隆表达及亲和层析纯化[J].2001,(16).
- [2]张文红,刘新平,王吉村,等.兔抗人NDR2高效价抗血清的制备、纯化及鉴定[J].2002,(8).
- [3]邓艳春,王立波,刘新平,等.P40蛋白在大肠杆菌内的表达及初步纯化[J].2001,(1).
- [4]Tomohiko Okuda,Hisato Kondoh.Identification of New Genes Ndr2 and Ndr3 Which Are Related to Ndr1/RTP/Drg1 but Show Distinct Tissue Specificity and Response to N-myc[J].1999,266(1).
- [5]Rechsteiner M,Rogers SW.PEST SEQUENCES AND REGULATION BY PROTEOLYSIS [Review][J].1996,21(7).
- [6]Zhou RH,Yutani C,Kato H,etc.Characterization of the human NDRG gene family: a newly identified member, NDRG4, is specifically expressed in brain and heart.[J].2001,73(1).
- [7]D, Zhou,K, Salnikow,M, Costa.Cap43, a novel gene specifically induced by Ni2+ compounds.[J].1998,58.
- [8]S K, Kurdistani,P, Arizti,C L, Reimer,etc.Inhibition of tumor cell growth by RTP/rit42 and its responsiveness to p53 and DNA damage.[J].1998,58.
- [9]Diesveld MP,van der Made AC,Trapman J,etc.A novel gene which is up-regulated during colon epithelial cell differentiation and down-regulated in colorectal neoplasms.[J].1997,77(1).
- [10]Tien-Min Lin,Chawnshang Chang.Cloning and characterization of TDD5, an androgen target gene that is differentially repressed by testosterone and dihydrotestosterone[J].1997,94(10).
- [11]邓艳春,药立波,刘新平,等.人脑内一含有ACP样结构域新基因的发现[J].2001,(1).