Difference between revisions of "Part:BBa K2615005:Design"

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===Source===
 
===Source===
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<p>
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The part is designed by OUC-China. The wild type Csy4 existed in Pseudomonas aeruginosa. In Pseudomonas aeruginosa,
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Science Museum No.105，College of Marine Life Sciences, Ocean University of China
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crRNA biogenesis requires the endoribonuclease Csy4, which binds and cleaves the repetitive sequence of the CRISPR transcript.
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</p>
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Biochemical assays and three co-crystal structures of wild-type and mutant Csy4/RNA complexes reveal a substrate positioning and cleavage mechanism in which a histidine deprotonates the ribosyl 20-hydroxyl pinned in place by a serine, leading to nucleophilic attack on the scissile phosphate.
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===References===
 
===References===
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<p>
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[http://science.sciencemag.org/content/329/5997/1355 Haurwitz R E, Jinek M, Wiedenheft B, et al. Sequence- and structure-specific RNA processing by a CRISPR endonuclease[J]. Science, 2010, 329(5997):1355-1358.]
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</p>

Latest revision as of 13:43, 11 September 2018


Csy4-Y176F, the No.3 member of Csy4 family.


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 377
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 93


Design Notes

We designed this part by point mutation. We changed the TAC(encoding Tyr) to TTT(encoding Phe) on the 176th site based on wild type Csy4.

Source

Science Museum No.105，College of Marine Life Sciences, Ocean University of China

References

[http://science.sciencemag.org/content/329/5997/1355 Haurwitz R E, Jinek M, Wiedenheft B, et al. Sequence- and structure-specific RNA processing by a CRISPR endonuclease[J]. Science, 2010, 329(5997):1355-1358.]