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Bacteriophage lambda: A temperate inducible phage and type species of the genus lambda-like viruses, in the family Siphoviridae. Its natural host is E. coli K12. Its Virion contains linear double-stranded DNA with single-stranded 12-base 5' sticky ends. The DNA circularizes on infection.

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

1Department of Biology, California Institute of Technology, 2Department of Bioengineering, California Institute of Technology, 3Synthetic Biology Center, Department of Bioengineering, Massachusetts Institute of Technology, 4School of Physics and Astronomy, University of Minnesota

JoVE 50762


 Biology

Identifying DNA Mutations in Purified Hematopoietic Stem/Progenitor Cells

1Greehey Children's Cancer Research Institute, UT Health Science Center at San Antonio, 2Department of Cellular and Structural Biology, UT Health Science Center at San Antonio, 3Department of Pathology, UT Health Science Center at San Antonio, 4Department of Microbiology, UT Health Science Center at San Antonio, 5Cancer Therapy and Research Center, UT Health Science Center at San Antonio

JoVE 50752


 Immunology and Infection

Antibody Binding Specificity for Kappa (Vκ) Light Chain-containing Human (IgM) Antibodies: Polysialic Acid (PSA) Attached to NCAM as a Case Study

1Department of Neurology, Mayo Clinic, 2Mayo Clinic Center for Multiple Sclerosis and Autoimmune Neurology, Mayo Clinic, 3Center for Regenerative Medicine, Neuroregeneration, Mayo Clinic, 4Division of Neonatal Medicine, Mayo Clinic, 5Department of Pediatric and Adolescent Medicine, Mayo Clinic

JoVE 54139


 Immunology and Infection

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-Canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

1Institute of Chemistry L 1, Department of Biocatalysis, Technical University of Berlin, 2Institute of Chemistry PC 14, Department of Bioenergetics, Technical University of Berlin, 3Institute of Chemistry TC 7, Department of Physical Chemistry/Molecular Material Sciences, Technical University of Berlin

Video Coming Soon

JoVE 57017


 JoVE In-Press

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

1Center for AIDS Health Disparities Research, Meharry Medical College, 2Department of Biochemistry and Cancer Biology, Meharry Medical College, 3School of Graduate Studies and Research, Meharry Medical College, 4Department of Microbiology and Immunology, Meharry Medical College, 5Tennessee Center for AIDS Research (TN-CFAR), Meharry Medical College

JoVE 54581


 Immunology and Infection

Recombineering and Gene Targeting

JoVE 5553

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 on stretches of similar or identical sequences. Together with gene targeting, which takes advantage of endogenous HR to alter an organism’s genome at a specific loci, HR-based cloning techniques have greatly improved the speed and efficacy of high-throughput genetic engineering.In this video, we introduce the principles of HR, as well as the basic components required to perform a recombineering experiment, including recombination-competent organisms and genomic libraries such as bacterial artificial chromosomes (BAC). We then walk through a protocol that uses recombineering to generate a gene-targeting vector that can ultimately be transfected into embryonic stem cells to generate a transgenic animal. Finally, several applications that highlight the utility and variety of recombineering techniques wi


 Genetics

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An Optimized Enrichment Technique for the Isolation of Arthrobacter Bacteriophage Species from Soil Sample Isolates

1Biology Department, University of the Sciences, 2Biology Department, Arcadia University, 3Biology Department, Immaculata University, 4Biology/Clinical Laboratory Science, Neumann University, 5Science Department, Cabrini College

JoVE 52781


 Environment

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Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

1Centro Cardiologico Monzino IRCCS, 2Cardiovascular Tissue Bank of Milan, Centro Cardiologico Monzino IRCCS, 3Department of Clinical Sciences and Community Health, Cardiovascular Section, University of Milan, 4Department of Cardiovascular Disease, Development and Innovation Cardiac Surgery Unit, Centro Cardiologico Monzino IRCCS

JoVE 55762


 Biochemistry

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Phage Phenomics: Physiological Approaches to Characterize Novel Viral Proteins

1Department of Biology, San Diego State University, 2Computational Science Research Center, San Diego State University, 3Bioinformatics and Medical Informatics Research Center, San Diego State University, 4Department of Mathematics and Statistics, San Diego State University, 5Department of Computer Science, San Diego State University, 6Mathematics and Computer Science Division, Argonne National Laboratory, 7SPARC Committee, Broad Institute

JoVE 52854


 Immunology and Infection

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