Difference between revisions of "Part:BBa K512002"

 
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===Usage and Biology===
 
===Usage and Biology===
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Bacteria protect their genome and remove foreign DNA elements through the use of the clustered, regularly interspaced short palindromic repeat (CRIPSR) system. Recently discovered, this defense mechanism can protect microorganisms against bacteriaphages. The CRISPR system comprises a transcribed locus of repeats and spacers that is activated and made functional via a family of CAS proteins. The CRISPR repeats are short DNA sequences that are almost identical in size and sequence. Between a pair of repeats is a spacer, which if complementary to a DNA element of the invading bacteriaphage, facilitates the destruction of that sequence from the bacterial cell. As such, the CRISPR/CAS system is a primitive biological defense mechanism, similar in many ways to the process of adaptive immunity in eukaryotic immune systems. Plasmids, which are commonly used in molecular biology are also the source of horizontal gene transfer in the wild and facilitate the spread of antibiotic resistant genes among microorganisms. These plasmids can contain unique DNA sequences that could be targeted by the CRISPR/CAS system.
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Proof of operation:
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[[Image:Cas3 function.jpg]]
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In the chart above, BL-D1 are the cells without IPTG, BL-D2 are with IPTG; and for BL-D3, we add IPTG 2 hours later. IPTG can induce CRISPR to target the plasmid. As we can see from the chart, with cas3 and CRISPR, the growth rate and optical density are decreased significantly  compare to the cells without IPTG.
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For complete and other results, please check the link [http://2011.igem.org/Team:USC/Project]
  
 
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Revision as of 02:54, 29 September 2011

K12 CRISPR with GFP target spacer

CRISPR

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 222
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 222
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 222
    Illegal BamHI site found at 344
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 222
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 222
    Illegal AgeI site found at 265
  • 1000
    COMPATIBLE WITH RFC[1000]