ArticleMolecular biotechnology2007
Time-course determination of plasmid content in eukaryotic and prokaryotic cells using real-time PCR.
Article in Molecular biotechnology, 2007. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 21 papers.
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Who cites it
21 citing papers in PubMed, 52 citations in OpenAlex.
- Expanded Functional Characterization and Optimization of Protein Expression in Treponema denticola Shuttle Plasmids.Molecular oral microbiology · 2026Article
- Phosphorylation of SNAP-25 at Ser187 is enhanced following its cleavage by Botulinum Neurotoxin Serotype A, promoting the dominant-negative effect of the resulting fragment.PLoS pathogens · 2025Article
- Base editing rescues acid α-glucosidase function in infantile-onset Pompe disease patient-derived cells.Molecular therapy. Nucleic acids · 2024Article
- A Real-Time Quantitative PCR Protocol for the Quantification of Plasmid Copy Number in Lactococcus lactis.Methods in molecular biology (Clifton, N.J.) · 2023Article
- A statistical framework for determination of minimal plasmid copy number required for transgene expression in mammalian cells.Bioelectrochemistry (Amsterdam, Netherlands) · 2021Article
- The Impact of IPTG Induction on Plasmid Stability and Heterologous Protein Expression byInternational journal of molecular sciences · 2020Article
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- Copy number variability of expression plasmids determined by cell sorting and Droplet Digital PCR.Microbial cell factories · 2016Article
- Development of a synthetic gene network to modulate gene expression by mechanical forces.Scientific reports · 2016Article
- Quantification of cellular and nuclear uptake rates of polymeric gene delivery nanoparticles and DNA plasmids via flow cytometry.Acta biomaterialia · 2016Article
- Retrohoming of a Mobile Group II Intron in Human Cells Suggests How Eukaryotes Limit Group II Intron Proliferation.PLoS genetics · 2015Article
- A model for the evolution of biological specificity: a cross-reacting DNA-binding protein causes plasmid incompatibility.Journal of bacteriology · 2014Article
- Development of an equine-tropic replication-competent lentivirus assay for equine infectious anemia virus-based lentiviral vectors.Human gene therapy methods · 2012Article
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- Quantifying plasmid copy number to investigate plasmid dosage effects associated with directed protein evolution.Methods in molecular biology (Clifton, N.J.) · 2012Article
- BglBrick vectors and datasheets: A synthetic biology platform for gene expression.Journal of biological engineering · 2011Article
- Site-directed integration of transgenes: transposons revisited using DNA-binding-domain technologies.Genetica · 2010Article
- Nonviral gene delivery to mesenchymal stem cells using cationic liposomes for gene and cell therapy.Journal of biomedicine & biotechnology · 2010Article
- Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes.Microbial cell factories · 2008Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A Real-Time PCR method was developed to monitor the plasmid copy number (PCN) in Escherichia coli and Chinese hamster ovary (CHO) cells. E. coli was transformed with plasmids containing a ColE1 or p15A origin of replication and CHO cells were transfected with a ColE1 derived plasmid used in DNA vaccination and carrying the green fluorescent protein (GFP) reporter gene. The procedure requires neither specific cell lysis nor DNA purification and can be performed in <30 min with dynamic ranges covering 0.9 pg-55 ng, and 5.0 pg-2.5 ng of plasmid DNA (pDNA) for E. coli and CHO cells, respectively. Analysis of PCN in E. coli batch cultures revealed that the maximum copy number per cell is attained in mid-exponential phase and that this number decreases on average 80% towards the end of cultivation for both types of plasmids. The plasmid content of CHO cells determined 24 h post-transfection was around 3 x 104 copies per cell although only 37% of the cells expressed GFP one day after transfection. The half-life of pDNA was 20 h and around 100 copies/cell were still detected 6 days after transfection.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.