Title |
A simplified counter-selection recombineering protocol for creating fluorescent protein reporter constructs directly from C. elegans fosmid genomic clones
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Published in |
BMC Biotechnology, January 2013
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DOI | 10.1186/1472-6750-13-1 |
Pubmed ID | |
Authors |
Nisha Hirani, Marcel Westenberg, Minaxi S Gami, Paul Davis, Ian A Hope, Colin T Dolphin |
Abstract |
Recombineering is a genetic engineering tool that enables facile modification of large episomal clones, e.g. BACs, fosmids. We have previously adapted this technology to generate, directly from fosmid-based genomic clones, fusion gene reporter constructs designed to investigate gene expression patterns in C. elegans. In our adaptation a rpsL-tet(A) positive/negative-selection cassette (RT-cassette) is first inserted and then, under negative selection, seamlessly replaced with the desired sequence. We report here on the generation and application of a resource comprising two sets of constructs designed to facilitate this particular recombineering approach. |
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