ΦC31-mediated cassette exchange into a bacterial artificial chromosome. (November 2007)

• Record Type: Journal Article
• Authors: Blaas, Leander; Musteanu, Monica; Zenz, Rainer; Eferl, Robert; Casanova, Emilio
• Is Part Of: Biotechniques. Volume 43:Number 5(2007)
• Page Start: 659
• Page End: 664
• Publication Date: 2007-11
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/000112592
• Languages: English
• ISSNs: 0736-6205

Abstract

The use of bacterial artificial chromosomes (BACs) modified via homologous recombination in Escherichia coli has become a powerful tool in the transgenic field. Homologous recombination allows the manipulation of BACs in very different ways. However, this process can be cumbersome and problematic when using large targeting constructs containing several repeated elements. In order to address this problem, we have established a ΦC31 integrase-mediated cassette exchange into a BAC. As an example of this technique, we have exchanged a cassette previously recombined into a BAC containing the Rosa 26 locus, by a 16.5-kb incoming construct containing several repeated elements. The combination of homologous recombination in E. coli and cassette exchange should expand the tools for manipulating BACs, thus facilitating the generation of constructs with higher complexity.