In vivo construction of recombinant molecules within the Caenorhabditis elegans germ line using short regions of terminal homology.

Abstract:

:Homologous recombination provides a means for the in vivo construction 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.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Kemp BJ,Hatzold J,Sternick LA,Cornman-Homonoff J,Whitaker JM,Tieu PJ,Lambie EJ

doi

10.1093/nar/gkm857

subject

Has Abstract

pub_date

2007-01-01 00:00:00

pages

e133

issue

19

eissn

0305-1048

issn

1362-4962

pii

gkm857

journal_volume

35

pub_type

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