Construction of malate-sensing <Emphasis Type="Italic">Escherichia coli</Emphasis> by introduction of a novel chimeric two-component system |
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Authors: | Irisappan Ganesh Sambandam Ravikumar Ik-keun Yoo Soon Ho Hong |
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Institution: | 1.Department of Chemical Engineering,University of Ulsan,Ulsan,Republic of Korea;2.Department of Pharmaceutical Science and Technology,Catholic University of Daegu,Gyeongsan-si,Republic of Korea |
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Abstract: | In an attempt to develop a high-throughput screening system for screening microorganisms which produce high amounts of malate, a MalKZ chimeric HK-based biosensor was constructed. Considering the sequence similarity among Escherichia coli (E. coli) MalK with Bacillus subtilis MalK and E. coli DcuS, the putative sensor domain of MalK was fused with the catalytic domain of EnvZ. The chimeric MalK/EnvZ TCS induced the ompC promoter through the cognate response regulator, OmpR, in response to extracellular malate. Real-time quantitative PCR and GFP fluorescence studies showed increased ompC gene expression and GFP fluorescence as malate concentration increased. By using this strategy, various chimeric TCS-based bacteria biosensors can be constructed, which may be used for the development of biochemical-producing recombinant microorganisms. |
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