Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites.

Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Research Abstract Details 

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Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Abstract Text:

Swetha Bolusani,Chien-Hui Ma,Andrew Paek,Jay H Konieczka,Makkuni Jayaram,Yuri Voziyanov,

As a tool in directed genome manipulations, site-specific recombination is a double-edged sword. Exquisite specificity, while highly desirable, makes it imperative that the target site be first inserted at the desired genomic locale before it can be manipulated. We describe a combination of computational and experimental strategies, based on the tyrosine recombinase Flp and its target site FRT, to overcome this impediment. We document the systematic evolution of Flp variants that can utilize, in a bacterial assay, two sites from the human interleukin 10 gene, IL10, as recombination substrates. Recombination competence on an end target site is acquired via chimeric sites containing mixed sequences from FRT and the genomic locus. This is the first time that a tyrosine site-specific recombinase has been coaxed successfully to perform DNA exchange within naturally occurring sequences derived from a foreign genomic context. We demonstrate the ability of an Flp variant to mediate integration of a reporter cassette in Escherichia coli via recombination at one of the IL10-derived sites.

Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Publishing Authors By Initials

S Bolusani,CH Ma,A Paek,JH Konieczka,M Jayaram,Y Voziyanov,

For similar fungi: yeasts research abstracts see: fungi: yeasts research

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Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Journal Published:

PUBLICATION TYPE: Research Support, U.S. Gov't,

Journal: Nucleic acids research

VOLUME: 34

Page Numbers: 5259-69

Journal Abbreviation: Nucleic Acids Res.

ISSN: 1362-4962

DAY: 26

MONTH: 09

YEAR: 2006

Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Information

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LANGUAGE: eng

NlmUniqueID: 411011

Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Keywords Mesh Terms:

KEYWORDS: Yeasts

MESH TERMS: genetics

Chemical & Substance for Abstract: Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites. Information

Substance Name: FLP recombinase

Registry Number: EC 2.7.7.-

Grant and Affiliation Information for Evolution of variants of yeast site-specific recombinase Flp that utilize native genomic sequences as recombination target sites.

AFFILIATION: School of Biological Sciences/IfM, 911 Hergot Avenue, Louisiana Tech University, Ruston, LA 71272, USA.

Country: England

AGENCY: United States NIGMS

GRANT: GM 35654

ACRONYM: GM

MEDLINETA: Nucleic Acids Res

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