Journal article
Using phiC31 integrase to make transgenic Xenopus laevis embryos
Nature protocols, Vol.1(3), pp.1248-1257
09/2006
DOI: 10.1038/nprot.2006.183
PMID: 17406408
Abstract
Bacteriophage phiC31 produces the enzyme integrase that allows the insertion of the phage genome into its bacterial host. This enzyme recognizes a specific DNA sequence in the phage (attP) and a different sequence in the bacterium (attB). Recombination between these sites leads to integration in a reaction that requires no accessory factors. Seminal studies by the Calos laboratory demonstrated that the phiC31 integrase was capable of integrating plasmid with an attB site into mammalian genomes at sites that approximated the attP site. We describe the use of attB-containing plasmids with insulated reporter genes for the successful integration of DNA into Xenopus embryos. The method offers a way to produce transgenic embryos without manipulation of sperm nuclei using microinjection methods that are standard for experiments in Xenopus laevis. The method aims to allow the non-mosaic controlled expression of new genetic material in the injected embryo and compares favorably with the time that is normally taken to analyze embryos injected with mRNAs, plasmids, morpholinos or oligonucleotides.
Details
- Title: Subtitle
- Using phiC31 integrase to make transgenic Xenopus laevis embryos
- Creators
- Daniel L Weeks - Department of Biochemistry, Bowen Science Building, University of IowaBryan G Allen - Department of Biochemistry, Bowen Science Building, University of Iowa
- Resource Type
- Journal article
- Publication Details
- Nature protocols, Vol.1(3), pp.1248-1257
- DOI
- 10.1038/nprot.2006.183
- PMID
- 17406408
- NLM abbreviation
- Nat Protoc
- ISSN
- 1754-2189
- eISSN
- 1750-2799
- Language
- English
- Date published
- 09/2006
- Academic Unit
- Stead Family Department of Pediatrics; Radiation Oncology; Biochemistry and Molecular Biology
- Record Identifier
- 9984024410502771
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