1. Natural LacI from E. coli yields faster response and higher level of expression than the LVA-tagged LacI.
- Author
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Andreassen PR, Fredberg S, Horan M, Knudsen MH, Jacobsen K, Kjær A, Krogh TJ, Kronborg T, Mattsson NC, Schmidt SI, Wille H, and Andersen A
- Subjects
- Escherichia coli metabolism, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Spectrometry, Fluorescence, Escherichia coli genetics, Genetic Engineering methods, Lac Operon genetics, Promoter Regions, Genetic genetics
- Abstract
The lac promoter is one of the most commonly used promoters for expression control of recombinant genes in E. coli. In the absence of galactosides, the lac promoter is repressed by its repressor protein LacI. Since the lac promoter is regulated by a repressor, overexpression of LacI is necessary for regulation when the promoter is introduced on a high-copy plasmid. For that purpose, a modified variant of LacI, a LVA-tagged LacI, was submitted to the Registry of Standard Biological Parts and has been used for more than 500 constructs since then. We have found, however, that natural LacI is superior to the LVA-tagged LacI as controller of expression.
- Published
- 2014
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