Lambda Red Recombination
Lambda Red Recombination - Related Videos
06:10
Following Cell-fate in E. coli After Infection by Phage Lambda
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Cited by 20 •
2011
This article describes the procedure for preparing a fluorescently-labeled version of bacteriophage lambda, infection of E. coli bacteria, following the infection outcome under the microscope, and analysis of infection results.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
07:08
The Lambda Select cII Mutation Detection System
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Cited by 3 •
2018
We describe a detailed protocol for the Lambda Select cII mutation assay in cultured cells of transgenic rodents or the corresponding animals treated with a chemical/physical agent of interest. This approach has been widely used for mutagenicity testing of carcinogens in mammalian cells.
14:23
Recombineering Homologous Recombination Constructs in Drosophila
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Cited by 7 •
2013
Homologous recombination techniques greatly advance Drosophila genetics by enabling the creation of molecularly precise mutations. The recent adoption of recombineering allows one to manipulate large pieces of DNA and transform them into Drosophila6. The methods presented here combine these techniques to rapidly generate large homologous recombination vectors.
12:03
The Production of C. elegans Transgenes via Recombineering with the galK Selectable Marker
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Cited by 4 •
2011
The ability to produce transgenes for Caenorhabditis elegans using genomic DNA carried by fosmids is particularly attractive as all of the native regulatory elements are retained. Described is a simple and robust procedure for the production of transgenes via recombineering with the galK selectable marker.
FAQs
Gam, Exo, and Beta divide the recombination task into complementary functions. Gam limits RecBCD-mediated destruction of linear donor DNA, Exo processes donor ends to expose single-stranded regions, and Beta supports pairing with matching chromosomal sequences. Their coordination links donor preservation, strand processing, and sequence alignment, allowing a designed DNA change to be incorporated at a selected bacterial locus.
Linear donor DNA is vulnerable to degradation by the bacterial RecBCD system. Gam counteracts this barrier, helping preserve the donor molecule long enough for Exo and Beta to act on it. This protection is mechanistically important because successful modification depends not only on sequence matching, but also on maintaining an intact donor template in the bacterial cell.
Homology regions provide sequence correspondence between the donor DNA and the target chromosome. Beta promotes annealing where these matching sequences meet, so the engineered segment is positioned relative to the intended bacterial locus rather than incorporated randomly. Researchers can therefore design replacements, insertions, or tagged versions whose placement is defined by the donor's matching chromosomal sequences.
The method supports several targeted design goals, including replacing an existing gene, inserting a regulatory element, and adding an epitope or fluorescent tag. These options let researchers alter either the DNA sequence itself or the way a gene can be monitored and regulated. The same general strategy therefore supports both genome engineering and analysis of gene or protein behavior.
In functional genomics, targeted sequence changes help connect a gene with its biological role. Gene replacement can test the consequences of removing or altering a sequence, while regulatory insertions and fluorescent tags can help examine gene regulation or protein function. By creating defined bacterial variants, the method supports comparisons between engineered strains and their unmodified counterparts.
Bacterial artificial chromosomes can carry large DNA constructs that researchers may need to alter at selected positions. Lambda Red recombination provides a flexible way to replace sequences or add regulatory and tagging elements within these constructs. This expands its value beyond routine chromosomal edits, supporting the preparation of engineered DNA resources for functional studies and strain construction.