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

doi:10.11938/cjmr20150101

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

Abstract

Abstract:

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 be b-lactamase.

Key words: NRSF/REST, ZnF2-8, NMR spectroscopy, b-lactamase, unexpectedly overexpress

CLC Number:

O482.53

ZHANG Yan，ZHAO Bin，YANG Zhong-zheng，SHEN Jie，HU Wei，LAN Wen-xian，WU Hou-ming，CAO Chun-yang. Methods for Identification and Characterization of Protein Unexpectedly Expressed in Escherichia Coli: A Case Study Involving b-Lactamase Observed during the Expression of Zinc Finger 2-8 of NRSF/REST[J]. Chinese Journal of Magnetic Resonance, 2015, 32(1): 1-11.

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