原核表达过程中非目标蛋白质识别与确认的方法:NRSF/REST蛋白功能结构域ZnF2-8原核表达过程中β-内酰胺酶的确认（英文）  

Methods for Identification and Characterization of Protein Unexpectedly Expressed in Escherichia Coli: A Case Study Involving β-Lactamase Observed during the Expression of Zinc Finger 2-8 of NRSF/REST

作　　者：张岩[1] 赵玢[1] 杨中正[1] 申杰[1] 胡伟[1] 蓝文贤[1] 吴厚铭[1] 曹春阳[1] 

机构地区：[1]中国科学院上海有机化学研究所,生命有机国家重点实验室,上海200032

出　　处：《波谱学杂志》2015年第1期1-11,共11页Chinese Journal of Magnetic Resonance

基　　金：The National Basic Research Program of China(2011CB966300);the National Natural Science Foundation of China(30970595,21472229 and 20921091)

摘　　要：大肠杆菌常被用来大量快速制备重组蛋白质.但是,在原核表达目标蛋白质时非目标蛋白质经常会意外表达.有时这些非目标蛋白质也非常有使用价值,但是最终确认这些非目标蛋白质的过程昂贵又及其耗时.基于此,该文发展了一个新的基于二维核磁共振波谱技术、X-单晶衍射技术、结合其他生物化学方法,确认在原核表达神经限制性沉默因子NRSF/REST蛋白(该蛋白能够特异性识别神经限制性沉默因子RE1ds DNA及神经限制性激活因子ds RNA,以调节神经元干细胞的发育)功能结构域Zn F2-8时非目标蛋白β-内酰胺酶(β-lactamase).Escherichia coli （E. coli） is often used to produce recombinant proteins rapidly with high yield. However, the bacteria expresses non-target proteins unexpectedly in many cases, some of which may later be proven useful. Full characterization of these unpredicted proteins are usually expensive and time-consuming. In this study, we used E. coli to express neuron- restrictive silencer factor/REl-silencing transcription factor （NRSF/REST） functional motif ZnF2-8, which is involved in the interaction of NRSF/REST with neuron-restrictive silencer element （NRSE/RE1） dsDNA or small non-coding dsRNA for neuron gene transcriptional repression or activation. Overexpression of a non-target protein was observed. Two-dimensional 1H-15N HSQC NMR spectroscopy, X-ray crystallography and other biochemical assays were used in combination to characterize the non-target protein to be β-lactamase.

关 键 词：NRSF/REST ZnF2—8 核磁共振(NMR) Β-内酰胺酶 意外表达 

分 类 号：O482.53[理学—固体物理]