人源CBX7功能结构域的重组表达与纯化  

Recombinant Expression, Purification of a Human CBX7 Functional Domain

作　　者：胡温温 ALTAF Simair 吕缜一 陈婷[1] 张云龙[1] 陆昌瑞 HU Wen-wen;ALTAF Simair;LU Zhen-yi;CHEN Ting;ZHANG Yun-long;LU Chang-rui(College of Chemistry, Chemical Engineering and Biotechnology,Donghua University, Shanghai 201620,China)

机构地区：[1]东华大学化学化工与生物工程学院,中国上海201620

出　　处：《生命科学研究》2019年第2期92-99,共8页Life Science Research

基　　金：国家自然科学基金资助项目(31300603);中央高校基本科研基金资助项目(15D110508;15D110527;13D110522;17D210502;17D310512;2232014D3-39);国家大学生创新项目(14T10501);上海高校特聘教授(东方学者)计划(2012-28);东华大学励志计划(18D210501);中国博士后科研基金特别资助项目(2015M571455)

摘　　要：CBX7(chromobox 7)在细胞寿命延长和癌症发生中起着重要的调控作用。然而,获取足量的全长CBX7蛋白并用于结构研究极为艰难。实验阐述了CBX7功能结构域CBX7(aa 145～227)重组表达载体构建的详细过程。同时,通过Ni-NTA亲和层析对融合目的蛋白进行了纯化,并利用SDS-PAGE及Western-blot进行了鉴定分析。纯化后的融合蛋白样品通过离子交换和分子筛层析进一步分离剩余杂质。本实验得到的高浓度CBX7蛋白可用于结晶,为后续CBX7的结构和功能研究奠定了基础。CBX7 regulates the cellular lifespan extension and cancer progression. However, it has proved difficult to obtain the full length CBX7 in sufficient quantities for structural studies. Here the process to recombinantly express the CBX7 functional domain CBX7 (aa 145~227) was described. The target recombinant protein was purified by Ni-NTA affinity chromatography and analyzed by SDS-PAGE and Western-blot. Remaining contaminants were further removed by ion-exchange and size-exclusion chromatography. Through this procedure, milligrams of the CBX7 protein could be available for crystal screen in future studies of CBX7 structure and functions.

关 键 词：人源CBX7 功能结构域 蛋白质表达 蛋白质纯化 肿瘤抑制 

分 类 号：Q518[生物学—生物化学] Q71