Difference between revisions of "Part:BBa K1956019"
 
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== IIT Madras 2016's Characterization ==
 
== IIT Madras 2016's Characterization ==
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<p>'''Trigger RNA''': A part of RNA molecule, which codes for [https://parts.igem.org/Part:BBa_E1010 RFP] protein.<br>
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'''Switch RNA''': It was designed to repress the translation process by masking the RBS placed upstream of GFP protein coding part. After interacting with the RNA molecules of RFP gene, <span class="riboson">RIBOSON</span> would allow the RBS to open up to bind to ribosomal machinery for protein synthesis to take place.</p>
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<p>We performed our <span class="riboson">RIBOSON</span> experiment in <i>E. coli</i> BL21(DE3). We transformed our cloned device [https://parts.igem.org/Part:BBa_K1956019 BBa_K1956019], which has <span class="riboson">RIBOSON<sub>rfp</sub></span> switch under [https://parts.igem.org/Part:BBa_J23100 J23100] constitutive promoter, upstream of [https://parts.igem.org/Part:BBa_E0040 GFP] coding part and RFP generator under [https://parts.igem.org/Part:BBa_R0011 lacI+pL] promoter.</p>
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<p> We grew the cells in LB broth media + 40mM Glucose. After, 16 hrs of growth, we made two types of secondary culture: 1. Cells + LB broth + 40mM Glucose and 2. Cells + LB broth + 1mM IPTG.</p>
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<p>Positive Control: Device [https://parts.igem.org/Part:BBa_K1956015 K1956015], which produces GFP and RFP proteins.<br>
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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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[[Image:Iitm_riboson_proof.png|480px]]
 
[[Image:Iitm_riboson_proof.png|480px]]
  

Latest revision as of 18:58, 22 October 2016


RIBOSON with plasmid control

RIBOSON with plasmid control

IIT Madras 2016's Characterization

Trigger RNA: A part of RNA molecule, which codes for RFP protein.
Switch RNA: It was designed to repress the translation process by masking the RBS placed upstream of GFP protein coding part. After interacting with the RNA molecules of RFP gene, RIBOSON would allow the RBS to open up to bind to ribosomal machinery for protein synthesis to take place.

We performed our RIBOSON experiment in E. coli BL21(DE3). We transformed our cloned device BBa_K1956019, which has RIBOSONrfp switch under J23100 constitutive promoter, upstream of GFP coding part and RFP generator under lacI+pL promoter.

We grew the cells in LB broth media + 40mM Glucose. After, 16 hrs of growth, we made two types of secondary culture: 1. Cells + LB broth + 40mM Glucose and 2. Cells + LB broth + 1mM IPTG.

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 RIBOSON device.

Iitm riboson 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 1844
    Illegal AgeI site found at 1956
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 842