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Team:Alberta/Project/Recombineering
From 2009.igem.org
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What is Recombineering?Recombineering refers to the strategic use of recombination in vivo in order to reach a defined goal. In the case of BioBytes, a method is required to target the final construct to insertion at a specific place on the E. coli chromosome. To do this successfully, three components must be taken into account: - There must be a system for targeting the construct to a specific site for insertion - Activation of the recombination system must be under experimenter control - It must be possible to select for and verify colonies in which the insertion was successful |
TargetingThe BioBytes team has chosen to use a recombination system from bacteriophage lambda. Lambda Red recombinase specifically recombines on the ends of a linear fragment of DNA. If the ends of this fragment are homologous to two separate sites on the E. coli chomosome, the genetic material between these two homologous regions will be exchanged. This will be the basis for our targeting system. According to literature on the topic, the homologous regions must be a minimum of 50 base pairs in length for recombination to occur a significant frequencies. |
