Difference between revisions of "Part:BBa K143001"

Revision as of 14:32, 18 September 2008

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.

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

1 pmid=3019840
2 pmid=14597697
3 Bacillus Genetic Stock Center [www.bgsc.org]
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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 __NOTOC__
-===<big>5' Integration Sequence for the AmyE locus of ''B. subtilis''</big>===
+===<big>5' Integration Sequence for the AmyE Locus of ''B. subtilis''</big>===
 [[Image:AmyE_Integration.png|center|800px]]
-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 (<bbpart>BBa_K143002</bbpart>) to the rear of the Biobrick construct to allow integration of the Biobrick construct into the chromosome of the gram positive bacterium ''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 (<bbpart>BBa_K143002</bbpart>) 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<cite>#1</cite> and is still commonly used in vectors such as pDR111<cite>#2</cite>, pDL<cite>#3</cite> 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<cite>#4</cite>. The 5' and 3' integration sequences for the AmyE locus were used to integrate the Imperial 2008 iGEM project primary construct into the ''B. subtilis'' chromosome.
+The AmyE locus was the first locus used for integration into ''B. subtilis'' by Shimotsu and Henner<cite>#1</cite> and is still commonly used in vectors such as pDR111<cite>#2</cite>, pDL<cite>#3</cite> 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<cite>#4</cite>. 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===