Using an in vivo phagemid system to identify non-compatible loxP sequences.

Abstract:

:The site-specific recombination system of bacteriophage P1 is composed of the Cre recombinase that recognizes a 34-bp loxP site. The Cre/loxP system has been extensively used to manipulate eukaryotic genomes for functional genomic investigations. The creation of additional heterologous loxP sequences potentially expands the utility of this system, but only if these loxP sequences do not recombine with one another. We have developed a stringent in vivo assay to examine the degree of recombination between all combinations of each previously published heterologous loxP sequence. As expected, homologous loxP sequences efficiently underwent Cre-mediated recombination. However, many of the heterologous loxP pairs were able to support recombination with rates varying from 5 to 100%. Some of these loxP sequences have previously been reported to be non-compatible with one another. Our study also confirmed other heterologous loxP pairs that had previously been shown to be non-compatible, as well as defined additional combinations that could be used in designing new recombination vectors.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Siegel RW,Jain R,Bradbury A

doi

10.1016/s0014-5793(01)02541-8

keywords:

subject

Has Abstract

pub_date

2001-06-15 00:00:00

pages

147-53

issue

1-2

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(01)02541-8

journal_volume

499

pub_type

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