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KMID : 0364820100460010104
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Korean Journal of Microbiology
2010 Volume.46 No. 1 p.104 ~ p.108
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Purification and Fluorometric Analysis of Leucine-Responsive Regulatory Protein from Escherichia coli
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Lee Chan-Yong
Kim Sung-Chul
Seo Cho-Hee
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Abstract
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We describe the construction of derivatives of wild type and mutant lrp genes that encode 6XHis-tag Lrps. These derivatives of wild type and mutant Lrp could be useful for in vitro studies including Lrp conformational changes. We show that 6XHis-tag Lrp wild type and 6XHis-tag Lrp R145W bind with similar patterns in vitro to 21 bp duplex DNA containing the consensus sequences of Lrp sites of upstream of the ilvIH operon. In addition, we report here the 6XHis-tag Lrp R145W is useful to investigate the conformational changes of Lrp in solution by using its own intrinsic fluorescence characteristics.
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KEYWORD
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global regulator, leucine, Leucine-responsive regulatory protein (Lrp)
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