Bacterial Artificial Chromosome Recombineering Core

The mission of the BAC Core is to provide access to a complex technology to investigators at the so that they can use their resources to conduct research instead of developing tools for research.

 BACs contain large segments of chromosomal DNA (over 100,000 base pairs). The extensive DNA sequence contained in the BACs confers significant advantages to investigators who wish to use gene expression in their research. Shorter DNA fragments often do not contain enough gene expression information to recapitulate normal gene expression patterns (Giraldo and Montoliu, 2001). The BAC Core uses recombineering methods  to introduce genetic modifications in BACs by homologous recombination.

The BAC Core
    purifies DNA from BAC clones for use in cell electroporation or the production of transgenic mice or rats
    BAC DNA purification methods are described here
    conducts pulsed field gel electrophoresis to restriction enzyme map BACs for purposes of identification
    introduces reporter proteins such as green fluorescent proteins to mark cells
    introduces point mutations in genes to modify the proteins produced in cells
    introduces deletions into genes in the BAC to modifiy proteins or abrogate expression
    knocks in cDNAs for expression for gene expression by heterlogous promoters
    subclones BAC DNA fragments with gap repair plasmids
    prepares gene targeting vectors for use in embryonic stem cells from BACs

BAC DNA Purification and pulsed field gel mapping request form.
BAC Recombineering request form.

Examples of research projects that take advantage of BAC technology include establishment of transgenic mouse models of human diseases (Cui et al., 2000), and transgenic mouse experiments to identify homologs of human disease genes (Probst et al., 1998, Wang et al., 1998). The Transgenic Core has a extensive experience in the production of BAC transgenic mice (Van Keuren et al., 2009) and transgenic rats so that the translation of genetically engineered BACs into genetically engineered mouse and rat models is straightforward.

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