Cell lysis in Escherichia coli cultures stimulates growth and biosynthesis of recombinant proteins in surviving cells

Corchero, José Luís; Cubarsi, Rafael; Vila, Pau; Arı́s, Anna; Villaverde, Antonio

doi:10.1078/0944-5013-00066

Cited by 20 publications

(15 citation statements)

References 25 publications

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“…This can easily be explained by the fact that inhibiting the growth of plasmid-free cells makes all resources available to plasmid-bearing bacteria. Moreover, cell lysis of the plasmid-free cells could even stimulate growth of surviving (plasmid-bearing) cells, as previously shown by Corchero and coworkers (28). …”

Section: Discussionmentioning

confidence: 66%

Separate-Component-Stabilization System for Protein and Dna Production Without the Use of Antibiotics

Szpirer

Milinkovitch

2005

BioTechniques

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Plasmid instability is a significant concern in the industrial utilization of microorganisms for protein or DNA production. Here we report on the development of a new and highly effective stabilization system based on the use of the ccd antidote/poison genes. For the first time, we separated the antidote gene from the poison gene: localizing the former in the plasmid and integrating the latter in the bacterial chromosome. We show that this separate-component-stabilization (SCS) strategy: (i) allows for perfect stabilization without the use of antibiotics; (ii) increases three to five times the recombinant protein production levels; and (iii) does not require any specific modification of the protein production process or culture medium. We illustrate that point by using the classical T7 promotor (i.e., used in most expression systems). Finally, we demonstrate that the SCS system increases by five the yield in DNA production, a result especially important for the design and production of gene therapy constructs void of any antibiotic resistance gene.

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Section: Discussionmentioning

confidence: 66%

Separate-Component-Stabilization System for Protein and Dna Production Without the Use of Antibiotics

Szpirer

Milinkovitch

2005

BioTechniques

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“…Long-term survival would be possible in fluid inclusions in salt deposits, and the maintenance energy might stem from scarifying a certain fraction of the enclosed population. For E. coli, it has indeed been shown that lysis of a fraction of a stationary-phase culture can even foster growth of the survivors [20].…”

Section: Survival Over Geological Timesmentioning

confidence: 99%

Evolutionary advantages of polyploidy in halophilic archaea

Soppa

2013

Biochemical Society Transactions

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Several species of haloarchaea have been shown to be polyploid and thus this trait might be typical for and widespread in haloarchaea. In the present paper, nine different possible evolutionary advantages of polyploidy for haloarchaea are discussed, including low mutation rate, radiation/desiccation resistance, gene redundancy and survival over geological times and at extraterrestrial sites. Experimental indications exist for all but one of these evolutionary advantages. Several of the advantages require gene conversion, which has been shown to be present and active in haloarchaea.

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“…The maintenance energy for long-term survival, including the repair of double-strand breaks, might originate from lysis of some or even the majority of cells of the enclosed population. For Escherichia coli, it has indeed been shown that cell lysis promotes growth of the remaining population (Corchero et al, 2001). Recently, three haloarchaeal species have been freshly isolated from an ancient salt deposit, and their ploidy levels have been quantified.…”

Section: Long-term Survival and Survival In Extreme Environmentsmentioning

confidence: 99%

Polyploidy in haloarchaea: advantages for growth and survival

Zerulla

Soppa

2014

Front. Microbiol.

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The investigated haloarchaeal species, Halobacterium salinarum, Haloferax mediterranei, and H. volcanii, have all been shown to be polyploid. They contain several replicons that have independent copy number regulation, and most have a higher copy number during exponential growth phase than in stationary phase. The possible evolutionary advantages of polyploidy for haloarchaea, most of which have experimental support for at least one species, are discussed. These advantages include a low mutation rate and high resistance toward X-ray irradiation and desiccation, which depend on homologous recombination. For H. volcanii, it has been shown that gene conversion operates in the absence of selection, which leads to the equalization of genome copies. On the other hand, selective forces might lead to heterozygous cells, which have been verified in the laboratory. Additional advantages of polyploidy are survival over geological times in halite deposits as well as at extreme conditions on earth and at simulated Mars conditions. Recently, it was found that H. volcanii uses genomic DNA as genetic material and as a storage polymer for phosphate. In the absence of phosphate, H. volcanii dramatically decreases its genome copy number, thereby enabling cell multiplication, but diminishing the genetic advantages of polyploidy. Stable storage of phosphate is proposed as an alternative driving force for the emergence of DNA in early evolution. Several additional potential advantages of polyploidy are discussed that have not been addressed experimentally for haloarchaea. An outlook summarizes selected current trends and possible future developments.

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Cell lysis in Escherichia coli cultures stimulates growth and biosynthesis of recombinant proteins in surviving cells

Cited by 20 publications

References 25 publications

Separate-Component-Stabilization System for Protein and Dna Production Without the Use of Antibiotics

Separate-Component-Stabilization System for Protein and Dna Production Without the Use of Antibiotics

Evolutionary advantages of polyploidy in halophilic archaea

Polyploidy in haloarchaea: advantages for growth and survival

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