Replication of prophage P1 is cell-cycle specific

Keasling, Jay D.; Palsson, Bernhard Ø.; Cooper, Stephen

doi:10.1128/jb.174.13.4457-4462.1992

Cited by 16 publications

(7 citation statements)

References 18 publications

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“…Consistent copy number in all cells of a culture can be achieved by timing the replication of the plasmid with the cell cycle, to ensure that the copy number has doubled before division (20)(21)(22)(23)(24), and by evenly partitioning plasmids into future daughter cells preceding division (25). Only the low-copy-number plasmids of E. coli (and, of course, the chromosome) have replication timed with the cell cycle (20)(21)(22)(23)(24).…”

Section: All-or-none Induction or Expressionmentioning

confidence: 99%

Synthetic Biology for Synthetic Chemistry

2008

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The richness and versatility of biological systems make them ideally suited to solve some of the world's most significant challenges, such as converting cheap, renewable resources into energy-rich molecules; producing high-quality, inexpensive drugs to fight disease; and remediating polluted sites. Over the years, significant strides have been made in engineering microorganisms to produce fuels, bulk chemicals, and valuable drugs from inexpensive starting materials; to detect and degrade nerve agents as well as less toxic organic pollutants; and to accumulate metals and reduce radionuclides. The components needed to engineer the chemistry inside a microbial cell are significantly different from those commonly used to overproduce pharmaceutical proteins. Synthetic biology has had and will continue to have a significant impact on the development of these components to engineer cellular metabolism and microbial chassis to host the chemistry. The ready availability of more well-characterized gene expression components and hosts for chemical synthesis, standards for the connection of these components to make larger functioning devices, computer-aided design software, and debugging tools for biological designs will decrease both the time and the support needed to construct these designs. Some of the most important tools for engineering bacterial metabolism and their use for production of the antimalarial drug artemisinin are reviewed.

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Section: All-or-none Induction or Expressionmentioning

confidence: 99%

Synthetic Biology for Synthetic Chemistry

2008

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“…Keasling et al [2,11] proposed that the F plasmid and P1 prophage replicated when the mass per plasmid origin reached a critical value (the initiation mass). This mechanism would be analogous to that proposed for the chromosomal origin [8,12].…”

Section: Model For the Timing Of Plasmid Replicationmentioning

confidence: 99%

Cycle-specific replication of chromosomal and F-plasmid origins

Cooper

1998

FEMS Microbiology Letters

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There have been various proposals for the pattern of F-plasmid replication during the division cycle. Here we show that the recent studies of Gordon et al. (Cell 90, 1113^1121, 1997 on the duplication and segregation of green fluorescent protein (GFP) labeled replication origins of the Escherichia coli chromosome and the F plasmid during the division cycle support the proposal that the F plasmid replicates with a cell-cycle-specific (artiocyclic) pattern. z

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“…Recently, we showed that the low-copy F plasmid and P1 prophage replicate in a cell-cycle-specific manner and that replication occurs when a constant mass per origin is achieved (4,5). In contrast, the high-copy ColEl-type plasmids replicate in a cell-cycle-independent manner (9,14).…”

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“…In light of the similarities in structure and function of the R6K plasmid and the low-copy F and P1 plasmids (large, conjugative, multiple origins of replication, DNA polymerase I independent), one might have expected the R6K plasmid to replicate in a cell-cycle-specific manner like these low-copy plasmids (4,5). However, if one considers the very high copy number of Replication of the mini-R6K plasmid pMF26 and the minichromosome pAL49 during the E. coli division cycle.…”

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Replication of the R6K plasmid during the Escherichia coli cell cycle

1992

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The cell-cycle replication pattern of the R6K plasmid has been investigated by using the membrane-elution technique to produce cells labeUled at different times during the division cycle and scintillation counting for quantitative analysis of radioactive plasmid DNA. The high-copy plasmid R6K replicates exponentially in a cell-cycle-independent manner. A mini-R6K plasmid deleted for the onra origin of replication also replicates, exponentially in a cell-cycle--independent manner. Plasmid R6K is a relatively large, high-copy, conjugative resistance plasmid with three replication origins (7). In vivo, replication initiates from orio about 50% of the time, from oril3 about 35% of the time, and from ori-y about 15% of the time (1). R6K is unique in that most large, self-transmissible plasmids exist at low copy number and are DNA polymerase I independent, whereas most high-copy plasmids are small, nonconjugative, and DNA polymerase I dependent (6).Recently, we showed that the low-copy F plasmid and P1 prophage replicate in a cell-cycle-specific manner and that replication occurs when a constant mass per origin is achieved (4, 5). In contrast, the high-copy ColEl-type plasmids replicate in a cell-cycle-independent manner (9, 14). Considering copy number alone, one might expect R6K to replicate in a cell-cycle-independent manner. Taking into account structural and functional characteristics of the plasmid, one might expect R6K to replicate in a cell-cyclespecific manner. The replication pattern has not been studied previously. In order to solve this dilemma, we have now investigated the replication patterns of the R6K plasmid and mini-R6K plasmid pMF26 (deleted for oria) by using the membrane-elution technique and quantitative determination of label incorporated into plasmid DNA.Escherichia coli B/r F65 (provided by Alan Leonard) containing the minichromosome pAL49 (7.6 kb, Kanr; provided by A. C. Leonard) and the R6K plasmid (Ampr Strr; provided by D. R. Helinski) or the mini-R6K plasmid pMF26 (Ampr; provided by D. R. Helinski) was grown for six to nine generations in 100 ml of C medium (3) supplemented with the amino acids histidine (42 ,ug/ml), proline (46 ,g/ml), and thymine (20 jig/ml) and with glucose (0.4%) or succinate (0.4%) at 37°C in order to obtain exponential growth prior to starting a membrane-elution experiment. The exponentially growing cells (100 ml, 108 cells per ml) were labelled with[3H]methyl thymidine (60 to 80 Ci/mmol, 10 RCi/ml) for 5% of the doubling time and filtered onto a nitrocellulose membrane at the end of the labelling period. The cells were washed with medium of the same composition containing cold thymidine (100 ,ug/ml). The membrane apparatus was inverted, and newborn cells were eluted from the membrane with prewarmed C medium pumped at a rate of 2 ml/min in * Corresponding author. t Present address: Department of Chemical Engineering, University of California, Berkeley, CA 94720. a 37°C incubator. Fractions were collected for an interval of time equivalent to the labelling period. Ce...

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Replication of prophage P1 is cell-cycle specific

Cited by 16 publications

References 18 publications

Synthetic Biology for Synthetic Chemistry

Synthetic Biology for Synthetic Chemistry

Cycle-specific replication of chromosomal and F-plasmid origins

Replication of the R6K plasmid during the Escherichia coli cell cycle

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