Emergent Properties of Reduced-Genome Escherichia coli

  title={Emergent Properties of Reduced-Genome Escherichia coli},
  author={Gy{\"o}rgy P{\'o}sfai and Guy Plunkett and Tam{\'a}s Feh{\'e}r and David A. Frisch and G{\"u}nther M. Keil and Kinga Umenhoffer and Vitaliy Kolisnychenko and Buffy Stahl and Shamik S Sharma and Monika de Arruda and Valerie Burland and Sarah W. Harcum and Frederick R. Blattner},
  pages={1044 - 1046}
With the use of synthetic biology, we reduced the Escherichia coli K-12 genome by making planned, precise deletions. The multiple-deletion series (MDS) strains, with genome reductions up to 15%, were designed by identifying nonessential genes and sequences for elimination, including recombinogenic or mobile DNA and cryptic virulence genes, while preserving good growth profiles and protein production. Genome reduction also led to unanticipated beneficial properties: high electroporation… 

Scarless engineering of the Escherichia coli genome.

Two genome engineering techniques, a lambda Red-mediated deletion method and a suicide (conditionally replicative) plasmid-based allele replacement procedure are presented here, and allow the rapid introduction of virtually any modifications in the genome.

Genetic manipulations restored the growth fitness of reduced-genome Escherichia coli.

Final Report Mechanisms of mutation in Escherichia coli : analysis , engineering and applications

  • Biology, Engineering
  • 2011
It is demonstrated that the lack of transposable elements aids in the cloning and expression of a heterologous protein, which is otherwise rapidly inactivated by insertion sequences disrupting its gene with a high frequency, and a method to measure the selective advantage conferred by a single IS element residing in the bacterial genome, compared to an IS-free host is developed.

Recombinant protein production in an Escherichia coli reduced genome strain.

Construction of Minimal Genomes and Synthetic Cells

This chapter will review previously constructed artificially reduced genomes to confirm the potential of their industrial utility and discuss some technological hurdles and limitations encountered during the design and construction of reduced genomes.

Directed homologous recombination for genome engineering in Escherichia coli.

Taking into account the findings at the assembly of the targeting plasmid constructs, serial genomic modifications can be created with enhanced efficiency and speed in the E. coli genome.

Superpositioning of Deletions Promotes Growth of Escherichia coli with a Reduced Genome

To retain robust growth and fundamental metabolic pathways, the growth of each deletion strain and combination effects of deletions were checked and a new strain, MGF-01, with a 1 Mb reduced genome was constructed by integrating deletions that did not affect growth.

Expression of two recombinant chloramphenicol acetyltransferase variants in highly reduced genome Escherichia coli strains

Use of a non‐catalytic CAT variant identified the recombinant protein activity as the source of the accumulation of acetate, and implications for the metabolic efficiency of the reduced genome strains are discussed.

Directed evolution of Escherichia coli with lower-than-natural plasmid mutation rates

A directed evolution strategy, Periodic Reselection for Evolutionarily Reliable Variants (PResERV), is developed to discover mutations that prolong the function of a burdensome DNA sequence in an engineered organism and promises to prevent evolutionary failures and unpredictability to provide a more stable genetic foundation for synthetic biology.

Enhancing recombinant protein production with an Escherichia coli host strain lacking insertion sequences

It is shown that IS hopping is detrimental to the industrial production of recombinant proteins, emphasizing the importance of the development of IS-free host strains.



Engineering a reduced Escherichia coli genome.

Techniques for precise genomic surgery were developed and applied to deleting the largest K-islands of E. coli, identified by comparative genomics as recent horizontal acquisitions to the genome, resulting in an 8.1% reduced genome size and reduction of gene count.

Minimization of the Escherichia coli genome using a Tn5-targeted Cre/loxP excision system

A genome-engineering tool for determining essential genes and minimizing bacterial genomes is developed and two large pools of independent transposon mutants in Escherichia coli are made using modified Tn5 transposons with two different selection markers.

The complete genome sequence of Escherichia coli K-12.

The 4,639,221-base pair sequence of Escherichia coli K-12 is presented and reveals ubiquitous as well as narrowly distributed gene families; many families of similar genes within E. coli are also evident.

Cell size and nucleoid organization of engineered Escherichia coli cells with a reduced genome

A simple system for formation of markerless chromosomal deletions is developed, and a series of large‐scale chromosomal deletion mutants of Escherichia coli that lack between 2.4 and 29.7% of the parental chromosome are constructed and characterized.

Mobile genetic elements: the agents of open source evolution

This review describes MGEs, their properties that are important in horizontal gene transfer, and current opportunities to advance MGE genomics.

Activation of the bgl operon by adaptive mutation.

  • B. Hall
  • Biology
    Molecular biology and evolution
  • 1998
In growing Escherichia coli K12 cells, the cryptic bgl operon is activated 98% of the time by insertions of IS1 or IS5 into the control region, designated bglR, providing the first direct evidence for adaptive mutagenesis mediated by the insertion of IS elements.

Differential plasmid rescue from transgenic mouse DNAs into Escherichia coli methylation-restriction mutants.

Statistical analysis of plasmid rescue frequencies has revealed that the MDRS loci detect differential modifications of the transgene insertions among mouse lines that show distinctive patterns of transgenes expression.