Difference between revisions of "Part:BBa K1956020"
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Negative Control: Device [https://parts.igem.org/Part:BBa_K1956034 K1956034], which produces lacI mRNA molecules.</p>
 
Negative Control: Device [https://parts.igem.org/Part:BBa_K1956034 K1956034], which produces lacI mRNA molecules.</p>
  
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Below result proves the functioning of RIBOSON device.
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Below result proves the functioning of RIBOSOFF device.
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[[Image:Iitm
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[[Image:Iitm_ribosoff_proof.png|480px]]
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K1956020 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K1956020 parameters</partinfo>
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Latest revision as of 19:07, 22 October 2016


RIBOSOFF with plasmid control

RIBOSOFF 1 with plasmid control

Team IIT Madras 2016's Characterization

Trigger RNA: A part of RNA molecule, which codes for LacI protein.
Switch RNA: It was designed to activate the translation process by allowing the ribosomal machinery to bind to RBS, placed upstream of GFP protein coding part. After interacting with the RNA molecules of lacI gene, RIBOSOFF changes it's confirmation to mask the RBS and represses the translation process.

We performed our RIBOSOFF experiment in E. coli DH5alpha. We transformed our cloned device BBa_K1956020, which has RIBOSOFFlacI switch under J23106 constitutive promoter, upstream of GFP coding part and RFP generator under lacI+pL promoter.

We prepared two types of cells: 1. BBa_K1956020 in pSB1A2 + BBa_K1956034 in pSB1K3, 2. BBa_K1956020 only.
We grew the cells in LB broth media. After, 16 hrs of growth, we inoculated secondary cultures and collect samples at 12 hour of growth.

Positive Control: Device K1956015, which produces GFP and RFP proteins.
Negative Control: Device K1956034, which produces lacI mRNA molecules.

Below result proves the functioning of RIBOSOFF device.

Iitm ribosoff proof.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1849
    Illegal AgeI site found at 1961
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 847