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AssigneeJiangnan University
InventorsLong LIU, Jian CHEN, Xueqin LV, Guocheng DU, Jianghua LI, Xianhao XU
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The present invention provides a pyruvate-responsive biosensor and a construction method and use thereof. In the present invention, pyruvate-responsive biosensors with different dynamic ranges are successfully constructed by optimizing the PdhR binding sequence inserted on the P43 promoter and optimizing the insertion site, wherein the minimum increase in dynamic range is by 0.6 time, and the maximum increase is by 30.7 times. The pyruvate-responsive biosensors are useful in the precise control of the expression of each gene in the cell. Since pyruvate is a key metabolite of central carbon in the cells, these biosensors are capable of dynamically regulating the expression level of intracellular genes according to changes in the content of pyruvate in the cells, thereby achieving the dynamic control of intracellular metabolic flux. The pyruvate biosensor obtained in the present invention has a good specificity, and a response range to pyruvate of 10-35 nmol/g DCW.
A pyruvate-responsive biosensor, comprising a PdhR gene, a P43 promoter, and a PdhR binding sequence inserted on the P43 promoter.
A phase-dependent pyruvate-responsive biosensor, comprising a PdhR gene, a phase-dependent promoter and a PdhR binding sequence inserted in the −35 region, the −10 region or the +1 site of the phase-dependent promoter, wherein the phase-dependent promoter is PyteJ, PywjC or Pyqfd.
Dependencies are inferred from the claim text.