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Human genomic site-specific recombination catalyzed by coliphage HK022 integrase

Paper ID Volume ID Publish Year Pages File Format Full-Text
24846 43541 2008 9 PDF Available
Title
Human genomic site-specific recombination catalyzed by coliphage HK022 integrase
Abstract

It has been previously demonstrated that the wild type integrase (Int) protein of coliphage HK022 can catalyze site-specific recombination in human cells between attachment (att) sites that were placed on extrachromosomal plasmids. In the present report it is shown that Int can catalyze the site-specific recombination reactions in a human cell culture on the chromosomal level. These include integrative (attP × attB) as well as excisive (attL × attR) reactions each in two configurations. In the cis configuration both sites are on the same chromosome, in the trans configuration one site is on a chromosome and the other on an episome. The reactions in cis were observed without any selection force, using the green fluorescent protein (GFP) as a reporter. The reactions in trans could be detected only when a selection force was applied, using the hygromycin-resistant (HygR) phenotype as a selective marker. All reactions were catalyzed without the need to supply any of the accessory proteins that are required by Int in its Escherichia coli host. The versatility of the att sites may be an advantage in the utilization of Int to integrate plasmid DNA into the genome, followed by a partial exclusion of the integrated plasmid.

Keywords
Amp, ampicillin; bp, base pairs; Ganc, Ganciclovir; GFP, green fluorescent protein; HygR, hygromycin resistant; IHF, integration host factor; Int, integrase; NeoR, neomycin resistant; CMV, cytomegalovirus promoter; RMCE, recombination mediated cassette ex
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Human genomic site-specific recombination catalyzed by coliphage HK022 integrase
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 134, Issues 1–2, 20 March 2008, Pages 46–54
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us