|COURT, DON - National Cancer Institute (NCI, NIH)|
Submitted to: Molecular Genetics of Bacteria and Phage
Publication Type: Abstract Only
Publication Acceptance Date: 7/10/2009
Publication Date: 8/4/2009
Citation: Swingle, B.M., Cartinhour, S.W., Court, D. 2009. Oligo Recombination in Gram Negative Bacteria. Molecular Genetics of Bacteria and Phage. p. 166.
Technical Abstract: Homologous recombination is important for bacterial survival because it simultaneously provides genomic stability as well as genomic plasticity. Of the mechanistic pathways for homologous recombination, those mediated by RecA are the most thoroughly characterized and are understood to be structurally and functionally conserved among all forms of life. In contrast, the phage-encoded recombinases mediate a distinct form of homologous recombination that is RecA-independent. The phage-encoded functions, such as lambda Red, have gained recognition for their ability to catalyze genomic changes using in vivo cloning strategies such as recombineering. Here we present results of a series of genetic experiments investigating the salient characteristics of a novel form of homologous recombination. The results show that oligonucleotides (oligos) can recombine with homologous genomic loci without the requirement for additional phage encoded recombinases. This form of oligo recombination is similar to lambda Red mediated recombination in several ways, including the lack of requirement for RecA and the replication imposed strand bias. However, oligo recombination differs from lambda Red mediated recombination with respect to tolerance to a wide range of oligo lengths and increased sensitivity to substrate oligo concentration. Additionally, we show that several species of Gram negative bacteria are capable of oligo recombination, suggesting that this process is evolutionarily conserved. This finding suggests that the ability of oligos to recombine with bacterial chromosomes may be applicable to a wide variety of species and useful for development of genetic systems in those organisms.