homologous recombination in vivo


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(PDF) Rapid cloning by homologous recombination in vi...

PDF | On Aug 1, 1993, Peter Bubeck and others published Rapid cloning by homologous recombination in vivo. We use cookies to make interactions with our website easy and meaningful, to better ...

In vitro recombination - Wikiped...

Recombinant DNA (rDNA), or molecular cloning, is the process by which a single gene, or segment of DNA, is isolated and amplified. Recombinant DNA is also known as in vitro recombination. A cloning vector is a DNA molecule that carries foreign DNA into a host cell, where it replicates, producing many copies of itself along with the foreign DNA ...

Homologous Recombination - an overview | ScienceDirect Topi...

Homologous recombination methods mimic the “sexual” recombination of genetic material that rearranges maternal and paternal chromosomes in germ cell DNA. Such recombination increases the genetic variation among a population and is vital to the continued evolution of organisms in response to an ever-changing environment [22].Unlike mutagenesis methods, which create novel diversity at the ...

Homologous Recombination Proteins and their Potential ....

Homologous Recombination Proteins 169 I. Introduction Homologous recombination is the natural means of targeting a gene for replacement by a homologue. By definition, the recombination process is spe­ cific, being limited to homologous target sites that exceed a species-imposed minimum length of DNA sequence similarity.

Harnessing homologous recombination in vitro generate ....

In vitro homologous recombination with and without RecA Homologous recombination in vivo depends upon a double-stranded break, generation of ssDNA by exonucleases, homology searching by recombinases, annealing of homologous stretches, and repair of overhangs and gaps by enzymes that include resolvases, nucleases and polymerases.

In vitro reconstitution of homologous recombination reactio...

In vitro reconstitution of homologous recombination reactions S. C. Kowalczykowski Division of Biological Sciences, Sections of Microbiology and of Molecular and Cellular Biology, University of California, Davis, Davis (California 95616, USA) Abstract.

Complex in vivo Ligation Using Homologous Recombination ....

The protocols presented here allow for the facile generation of a wide variety of complex multipart DNA constructs (tagged gene products, gene fusions, chimeric proteins, and other variants) using homologous recombination and in vivo ligation in budding yeast (Saccharomyces cerevisiae). This method is straightforward, efficient and cost-effective, and can be used both for vector creation and ...

In vivo Recombination After Chronic Damage Exposure Falls ....

In vivo Recombination After Chronic Damage Exposure Falls to Below Spontaneous Levels in ‘.pdf .Hendricks,2ScottCassie,1AndrewJ.Engelward,3andBevinP ...

Recombination Cloning in Bacter...

Recombination Cloning in Bacteria The following approach to high throughput subcloning using recombination in bacteria was developed in the Finley lab at Wayne State University by Jodi Parrish and Tom Limjindaporn (1). Introduction Co-transformation of a recA+ E. coli strain with a linearized plasmid and a linear fragment of DNA having

Homologous recombination occurs frequently at innate GT ....

In somatic cells, homologous recombination (HR) is a rare event caused by eventual DNA double-strand breaks (DSBs). In contrast, germ cells show high frequency of HR caused by programmed DSBs. Microsatellites are prone to DSBs during genome replication and, thereby, capable of promoting HR. It remains unclear whether HR occurs frequently at microsatellites both in normal somatic cells and germ ...

Recombination-mediated PCR-directed plasmid construction ....

Table 1 shows that recombination-mediated PCR cloning requires that the plasmid be linearized, as no recombinants were obtained from uncut plasmid (frequency <0.25%; see Table 1 legend for experimental details). The length of the homologous region also has a significant effect on recombination frequency.

Red/ET Recombination - genebridges.c...

Why is Red/ET Recombination a superior approach to DNA engineering? Red/ET Recombination allows a faster, more flexible and highly reliable modification of plasmids, BACs, or the E. coli genome than conventional cloning methods. Red/ET exploits phage l homologous recombination potential for in vivo genetic engineering in E. coli. Since Red/ET ...

Rapid cloning by homologous recombination in vi...

NucleicAcids Research, 1993, Vol. 21, No. 15 3601-3602 Rapid cloning by homologous recombination in vivo Peter Bubeck, Monica Winkler and Wilfried Bautsch* Institute of Medical Microbiology ...

Recombineering: Genetic Engineering in UNIT 1.16 Bacteria ....

Bacteria Using Homologous Recombination 1.16.2 Supplement 78 Current Protocols in Molecular Biology of steps is a recombineering reaction that replaces the sequence to be modified with an antibiotic-resistance cassette and a counter-selectable marker (e.g., sacB, which is toxic when cells are grown on medium containing sucrose; Gay et al., 1985).

Optimal Cloning of PCR Fragments by Homologous ....

PCR fragments and linear vectors containing overlapping ends are easily assembled into a propagative plasmid by homologous recombination in Escherichia coli. Although this gap-repair cloning approach is straightforward, its existence is virtually unknown to most molecular biologists. To popularize this method, we tested critical parameters influencing the efficiency of PCR fragments cloning ...

Homologous recombination - Wikiped...

Homologous recombination is a type of genetic recombination in which nucleotide sequences are exchanged between two similar or identical molecules of DNA. It is most widely used by cells to accurately repair harmful breaks that occur on both strands of DNA, known as double-strand breaks (DSB). Homologous recombination also produces new ...

Harnessing homologous recombination in vitro to generate ....

The third, MAGIC, is an in vivo method that relies upon homologous recombination and bacterial mating 15. These methods offer a uniform and seamless transfer of genes from one expression context ...

Shuffled antibody libraries created by in vivo homologous ....

Homologous recombination in yeast can be exploited to reliably generate libraries of >10 7 transformants from a pool of PCR products and a linearized plasmid vector. Homology in the PCR insertion products drives shuffling of these genes in vivo by yeast homologous recombination. Two scFvs that share 89.8% homology were shuffled in vivo by homologous recombination, and chimeric genes were ...

Rapid cloning by homologous recombination in viv...

Full text Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (247K), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Genetic Manipulation of Homologous Recombination In Vivo ....

Loss-of-function mutations in BLM , a member of the RecQ helicase family, cause Bloom's syndrome (BS), a rare, recessive genetic disorder that predisposes to many types of cancer. BLM functions in many aspects of DNA homeostasis, including the suppression of homologous recombination (HR) in somatic cells.