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...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 beβ-lactamase.展开更多
基金The National Basic Research Program of China(2011CB966300)the National Natural Science Foundation of China(30970595,21472229 and 20921091)
文摘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 beβ-lactamase.
