DNA
amyE 3 IS

Part:BBa_K143002

Designed by: Chris Hirst   Group: iGEM08_Imperial_College   (2008-08-28)
Revision as of 16:37, 18 September 2008 by Qiqin (Talk | contribs)

3' Integration Sequence for the AmyE locus of B. subtilis

AmyE Integration.png

Integration sequences allow DNA to be incorporated into the chromosome of a host cell at a specific locus 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_K143001) to the rear of the BioBrick construct to allow integration of the BioBrick construct into the chromosome of the Gram-positive bacterium B. subtilis.

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 iGEM 2008 competition, the Imperial Team used the 5' and 3' integration sequences for the AmyE locus to integrate genetic construct into the B. subtilis chromosome.

References

<biblio>

  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.

</biblio>


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Categories
//chassis/prokaryote/bsubtilis
//DNA/chromosomalintegration
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