ArticleProceedings of the National Academy of Sciences of the United States of America2009
Cloning human herpes virus 6A genome into bacterial artificial chromosomes and study of DNA replication intermediates.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers.
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Who cites it
22 citing papers in PubMed, 35 citations in OpenAlex.
- Development of viral infectious clones and their applications based on yeast and bacterial artificial chromosome platforms.Molecular biomedicine · 2025Review
- The emerging role of recombineering in microbiology.Engineering microbiology · 2023Review
- Direct cloning of a herpesvirus genome for rapid generation of infectious BAC clones.Journal of advanced research · 2023Article
- Article
- Methods for the Manipulation of Herpesvirus Genome and the Application to Marek's Disease Virus Research.Microorganisms · 2021Review
- Analysis of the Replication Mechanisms of the Human Papillomavirus Genomes.Frontiers in microbiology · 2021Article
- Repair of an Attenuated Low-Passage Murine Cytomegalovirus Bacterial Artificial Chromosome Identifies a Novel Spliced Gene Essential for Salivary Gland Tropism.Journal of virology · 2020Article
- Endogenization and excision of human herpesvirus 6 in human genomes.PLoS genetics · 2020Article
- Copy Number Heterogeneity, Large Origin Tandem Repeats, and Interspecies Recombination in Human Herpesvirus 6A (HHV-6A) and HHV-6B Reference Strains.Journal of virology · 2018Article
- Chromosomal integration of HHV-6A during non-productive viral infection.Scientific reports · 2017Article
- Highly Efficient CRISPR/Cas9-Mediated Homologous Recombination Promotes the Rapid Generation of Bacterial Artificial Chromosomes of Pseudorabies Virus.Frontiers in microbiology · 2016Article
- Rolling circle replication requires single-stranded DNA binding protein to avoid termination and production of double-stranded DNA.Nucleic acids research · 2014Article
- Article
- Reactivation of chromosomally integrated human herpesvirus-6 by telomeric circle formation.PLoS genetics · 2013Article
- A herpes simplex virus 2 glycoprotein D mutant generated by bacterial artificial chromosome mutagenesis is severely impaired for infecting neuronal cells and infects only Vero cells expressing exogenous HVEM.Journal of virology · 2012Article
- Small RNA deep sequencing identifies microRNAs and other small noncoding RNAs from human herpesvirus 6B.Journal of virology · 2012Article
- Human cytomegalovirus: bacterial artificial chromosome (BAC) cloning and genetic manipulation.Current protocols in microbiology · 2012Article
- Viral bacterial artificial chromosomes: generation, mutagenesis, and removal of mini-F sequences.Journal of biomedicine & biotechnology · 2012Review
- The molecular biology of human herpesvirus-6 latency and telomere integration.Microbes and infection · 2011Review
- Recent advances in cloning herpesviral genomes as infectious bacterial artificial chromosomes.Cell cycle (Georgetown, Tex.) · 2011Review
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Cloning of large viral genomes into bacterial artificial chromosomes (BACs) facilitates analyses of viral functions and molecular mutagenesis. Previous derivations of viral BACs involved laborious recombinations within infected cells. We describe a single-step production of viral BACs by direct cloning of unit length genomes, derived from circular or head-to-tail concatemeric DNA replication intermediates. The BAC cloning is independent of intracellular recombinations and DNA packaging constraints. We introduced the 160-kb human herpes virus 6A (HHV-6A) genome into BACs by digesting the viral DNA replicative intermediates with the Sfil enzyme that cleaves the viral genome in a single site. The recombinant BACs contained also the puromycin selection gene, GFP, and LoxP sites flanking the BAC sequences. The HHV-6A-BAC vectors were retained stably in puromycin selected 293T cells. In the presence of irradiated helper virus, supplying most likely proteins enhancing gene expression they expressed early and late genes in SupT1 T cells. The method is especially attractive for viruses that replicate inefficiently and for viruses propagated in suspension cells. We have used the fact that the BAC cloning "freezes" the viral DNA replication intermediates to analyze their structure. The results revealed that HHV-6A-BACs contained a single direct repeat (DR) rather than a DR-DR sequence, predicted to arise by circularization of parental genomes with a DR at each terminus. HHV-6A DNA molecules prepared from the infected cells also contained DNA molecules with a single DR. Such forms were not previously described for HHV-6 DNA.
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