Natural LacI from E. coli yields faster response and higher level of expression than the LVA-tagged LacI

ACS Synth Biol. 2014 Dec 19;3(12):949-52. doi: 10.1021/sb500031h.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Escherichia coli / genetics*
  • Escherichia coli / metabolism
  • Genetic Engineering / methods*
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Lac Operon / genetics*
  • Promoter Regions, Genetic / genetics*
  • Spectrometry, Fluorescence

Substances

  • Green Fluorescent Proteins