ArticleInternational journal of biochemistry and molecular biology2011
A multi-step strategy for BAC recombineering of large DNA fragments.
Article in International journal of biochemistry and molecular biology, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 7 papers.
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Who cites it
7 citing papers in PubMed, 8 citations in OpenAlex.
- Polymorphic tandem DNA repeats activate the human telomerase reverse transcriptase gene.Proceedings of the National Academy of Sciences of the United States of America · 2021Article
- Crispr/Cas9-mediated cleavages facilitate homologous recombination during genetic engineering of a large chromosomal region.Biotechnology and bioengineering · 2020Article
- ETS variant transcription factor 5 and c-Myc cooperate in derepressing the human telomerase gene promoter via composite ETS/E-box motifs.The Journal of biological chemistry · 2020Article
- Engineering a humanized telomerase reverse transcriptase gene in mouse embryonic stem cells.Scientific reports · 2019Article
- Regulation of human and mouse telomerase genes by genomic contexts and transcription factors during embryonic stem cell differentiation.Scientific reports · 2017Article
- Repression of telomerase gene promoter requires human-specific genomic context and is mediated by multiple HDAC1-containing corepressor complexes.FASEB journal : official publication of the Federation of American Societies for Experimental Biology · 2017Article
- Developments in the tools and methodologies of synthetic biology.Frontiers in bioengineering and biotechnology · 2014Review
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Authors and funding
3 authors at 1 institution in 1 country.
Funding
Abstract
Recombineering techniques have been developed to modify bacterial artificial chromosomes (BACs) via bacterial homologous recombination systems, simplifying the molecular manipulations of large DNA constructs. However, precise modifications of a DNA fragment larger than 2-3 kb by recombineering remain a difficult task, due to technical limitations in PCR amplification and purification of large DNA fragments. Here, we describe a new recombineering strategy for the replacement of large DNA fragments using the commonly utilized phage/Red recombination host system. This approach involved the introduction of rare restriction enzyme sites and positive selection markers into the ends of a large DNA fragment, followed by its release from the donor BAC construct and integration into an acceptor BAC. We have successfully employed this method to precisely swap a number of large DNA fragments ranging from 6 to 40 kb between two BAC constructs. Our results demonstrated that this new strategy was highly effective in the manipulations of large genomic DNA fragments and therefore should advance the conventional BAC recombineering technology to the next level.
Indexed as
Identifiers
22003432PMC3193292W184238261What OpenQuestion holds
Registered trials
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.