CAJ | 학술논문

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 RE1 dsDNA及神经限制性激活因子dsRNA，以调节神经元干细胞的发育）功能结构域ZnF2-8时非目标蛋白b-内酰胺酶(b-lactamase)。
대장간균상피용래대량쾌속제비중조단백질。단시，재원핵표체목표단백질시비목표단백질경상회의외표체。유시저사비목표단백질야비상유사용개치，단시최종학인저사비목표단백질적과정앙귀우급기모시。기우차，해문발전료일개신적기우이유핵자공진파보기술、X-단정연사기술、결합기타생물화학방법，학인재원핵표체신경한제성침묵인자 NRSF/REST 단백（해단백능구특이성식별신경한제성침묵인자 RE1 dsDNA급신경한제성격활인자dsRNA，이조절신경원간세포적발육）공능결구역ZnF2-8시비목표단백b-내선알매(b-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/RE1-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 beb-lactamase.