ArticleNucleic acids research2010
Selection of bacteriophage lambda integrases with altered recombination specificity by in vitro compartmentalization.
Article in Nucleic acids research, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed.
- Engineering cell-free systems by chemoproteomic-assisted phenotypic screening.RSC chemical biology · 2024Article
- Expanding the DNA editing toolbox: Novel lambda integrase variants targeting microalgal and human genome sequences.PloS one · 2024Article
- Characterization and Genomic Analysis of Marinobacter Phage vB_MalS-PS3, Representing a New Lambda-Like Temperate Siphoviral Genus Infecting Algae-Associated Bacteria.Frontiers in microbiology · 2021Article
- HK022 bacteriophage Integrase mediated RMCE as a potential tool for human gene therapy.Nucleic acids research · 2020Article
- Genome targeting by hybrid Flp-TAL recombinases.Scientific reports · 2020Article
- A novel λ integrase-mediated seamless vector transgenesis platform for therapeutic protein expression.Nucleic acids research · 2018Article
- High-throughput screening of biomolecules using cell-free gene expression systems.Synthetic biology (Oxford, England) · 2018Review
- High Throughput Screening and Selection Methods for Directed Enzyme Evolution.Industrial & engineering chemistry research · 2015Review
- Hybrid lentivirus-phiC31-int-NLS vector allows site-specific recombination in murine and human cells but induces DNA damage.PloS one · 2014Article
- Expanding the scope of site-specific recombinases for genetic and metabolic engineering.Biotechnology and bioengineering · 2014Review
- A Concept for Selection of Codon-Suppressor tRNAs Based on Read-Through Ribosome Display in an In Vitro Compartmentalized Cell-Free Translation System.Journal of nucleic acids · 2012Article
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4 authors.
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Abstract
In vitro compartmentalization (IVC) was employed for the first time to select for novel bacteriophage lambda integrase variants displaying significantly enhanced recombination activity on a non-cognate target DNA sequence. These variants displayed up to 9-fold increased recombination activity over the parental enzyme, and one mutant recombined the chosen non-cognate substrate more efficiently than the parental enzyme recombined the wild-type DNA substrate. The in vitro specificity phenotype extended to the intracellular recombination of episomal vectors in HEK293 cells. Surprisingly, mutations conferring the strongest phenotype do not occur in the lambda integrase core-binding domain, which is known to interact directly with cognate target sequences. Instead, they locate to the N-terminal domain which allosterically modulates integrase activity, highlighting a previously unknown role for this domain in directing integrase specificity. The method we describe provides a robust, completely in vitro platform for the development of novel integrase reagent tools for in vitro DNA manipulation and other biotechnological applications.
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