Part:BBa_K143001

5' Integration Sequence for the AmyE Locus of B. subtilis

Integration sequences allow DNA to be incorporated into the chromosome of a host cell at a specific locus. This is achieved by using leading (5') and trailing (3') DNA sequences that are the same as those at a specific locus of the chromosome. The 5' integration sequence can be added to the front of a BioBrick construct and the 3' integration sequence specific for this locus (BBa_K143002) to the rear of the Biobrick construct to allow integration of the BioBrick construct into the chromosome of the Gram-positive bacterium B. subtilis at the AmyE locus.

The AmyE locus was the first locus used for integration into B. subtilis by Shimotsu and Henner#1 and is still commonly used in vectors such as pDR111#2, pDL#3 and their derivatives. Integration at the AmyE locus removes the ability of B. subtilis to break down starch, which can be assayed with iodine as described by Cutting and Vander-horn#4. For the 2008 iGEM competition, the Imperial Team used the 5' and 3' integration sequences for the AmyE locus to integrate genetic constructs into the B. subtilis chromosome.

References

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1. 1 pmid=3019840
2. 2 pmid=14597697
3. 3 Bacillus Genetic Stock Center [www.bgsc.org]
4. 4 Cutting, S M.; Vander-Horn, P B. Genetic analysis. In: Harwood C R, Cutting S M. , editors. Molecular biological methods for Bacillus. Chichester, England: John Wiley & Sons, Ltd.; 1990. pp. 27–74.

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