Document Type
Article
Publication Date
9-26-2007
Publication Title
Nucleic Acids Research
Department
Department of Biological Sciences
Abstract
Homologous recombination provides a means for the in vivoconstruction of recombinant DNA molecules that may be problematic to assemble in vitro . We have investigated the efficiency of recombination within the Caenorhabditis elegans germ line as a function of the length of homology between recombining molecules. Our findings indicate that recombination can occur between molecules that share only 10 bp of terminal homology, and that 25 bp is sufficient to mediate relatively high levels of recombination. Recombination occurs with lower efficiency when the location of the homologous segment is subterminal or internal. As in yeast, recombination can also be mediated by either single- or double-stranded bridging oligonucleotides. We find that ligation between cohesive ends is highly efficient and does not require that the ends be phosphorylated; furthermore, precise intermolecular ligation between injected molecules that have blunt ends can also occur within the germ line.
DOI
10.1093/nar/gkm857
Original Citation
Kemp BJ, Hatzold J, Sternick LA, Cornman-Homonoff J, Whitaker JM, Tieu PJ, Lambie EJ. In vivo construction of recombinant molecules within the Caenorhabditis elegans germ line using short regions of terminal homology. Nucleic Acids Res. 2007;35(19):e133. doi: 10.1093/nar/gkm857. Epub 2007 Oct 11. PMID: 17933760; PMCID: PMC2095804.
Dartmouth Digital Commons Citation
Kemp, Benedict J.; Hatzold, Julia; Sternick, Laura A.; Cornman-Homonoff, Joshua; Whitaker, Jessica M.; Tieu, Pamela J.; and Lambie, Eric J., "In Vivo Construction of Recombinant Molecules within the Caenorhabditis Elegans Germ Line Using Short Regions of Terminal Homology" (2007). Dartmouth Scholarship. 3861.
https://digitalcommons.dartmouth.edu/facoa/3861