Difference between revisions of "Part:BBa K143001"

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===<big>5' Integration Sequence for the AmyE locus of ''B. subtilis''</big>===
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===<big>5' Integration Sequence for the amyE Locus of ''B. subtilis''</big>===
  
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[[Image:AmyE_Integration.png|center|800px]]
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[[Image:IntegrationAmyE.PNG|center|800px]]
  
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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''.  
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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'' at the amyE locus.
  
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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.
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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.
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<span class='h3bb'><big>'''Sequence and Features'''</big></span>
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<partinfo>BBa_K143001 SequenceAndFeatures</partinfo>
  
 
===References===
 
===References===

Latest revision as of 01:42, 30 October 2008

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

IntegrationAmyE.PNG

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.


Sequence and Features



Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

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>