Antonie van Leeuwenhoek

, Volume 95, Issue 4, pp 305–310

Induction of cadBA in an Escherichia coli lysine auxotroph transformed with a cad-gfp transcriptional fusion

Original Paper

DOI: 10.1007/s10482-009-9314-y

Cite this article as:
Chalova, V.I., Woodward, C.L. & Ricke, S.C. Antonie van Leeuwenhoek (2009) 95: 305. doi:10.1007/s10482-009-9314-y
  • 117 Downloads

Abstract

CadBA functions as a part of overall Escherichia coli response to low extracellular pH. A gfpmut3 structural gene transcriptionally fused to the cadBA promoter (Pcad) was used as a reporter to monitor changes in intracellular lysine as a potential factor influencing cadBA induction. Different patterns of cadBA induction were observed in two E. coli strains with different lysine biosynthetic capabilities. In E. coli ZK126 (pJBA25-Pcad), a lysine prototroph, maximum levels of induction were detected 3 h after the transfer of bacterial cells under inducing conditions (pH 5.8; 3.4 μM extracellular lysine). The induction subsequently decreased until hour 7 after which no further change in expression was observed. However, in the lysine depleted strain E. coli ATCC 23812 (pJBA25-Pcad) which is an auxotroph for lysine, no decrease in cadBA expression was observed over time under the same induction conditions. Although no time dependent statistical differences in intracellular lysine were observed, bacterial cells depleted for no longer than 4 h (1.38 ± 0.25 μmol lysine/g cell dry weight) exhibited more rapid induction of cadBA (after 3 h) and a lower maximum level of induction compared to cells with relatively lower intracellular lysine (approximately 1.08 μmol/g cell dry weight). For the latter, the detectable level of induction was delayed for 1 h but the maximum level of induction response was higher.

Keywords

Escherichia coli Lysine auxotroph cad operon Green fluorescent protein Transcriptional fusion Intracellular lysine 

Copyright information

© Springer Science+Business Media B.V. 2009

Authors and Affiliations

  • V. I. Chalova
    • 1
    • 2
  • C. L. Woodward
    • 1
  • S. C. Ricke
    • 1
    • 2
    • 3
  1. 1.Department of Poultry ScienceTexas A&M UniversityCollege StationUSA
  2. 2.Center for Food Safety, Department of Food Science, and Department of Poultry ScienceUniversity of ArkansasFayettevilleUSA
  3. 3.Department of Food ScienceUniversity of ArkansasFayettevilleUSA

Personalised recommendations