Difference between revisions of "Part:BBa K864440"
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<partinfo>BBa_K864440 short</partinfo>
 
<partinfo>BBa_K864440 short</partinfo>
  
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The natural small RNA spot42 taken from the E-coli MG1655. Small RNA is belived to interact in many metabolic pathways. The spot42 scaffold interacts with HFQ-proteins. It is in many cases shown to bind into the rbs area of the mRNA and thereby blocking translation initiation. ( V.Sharma, A.Yamamura Y.Yokobayashi, 2011 )
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This part is a component of the Uppsala University iGEM 2012 modular small RNA screening system, together with the target construct <partinfo>K864444</partinfo>.  
  
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We wanted to use this sRNA for gene downregulation on translational level, in this case block the ribosome from binding to the rbs area.  
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This is the small RNA spot42 from E coli MG1655 coupled to the strong promoter <partinfo>J23101</partinfo>. The spot42 regulates the galactose metabolism by binding to the mRNA and recruiting Hfq proteins [1], blocking initiation of translation. The spot42 has proven to be an especially suitable sRNA for engineering of silencing sRNA:s, by replacing its RNA-binding domain[2].  
  
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You can use this natural scaffold to make your own sRNA, inhibiting translation on a gene of your choice.
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To create a random sRNA library, the RNA-binding domain can be randomized by site-directed mutagenesis using the primers <partinfo>K864445</partinfo> and <partinfo>K864446</partinfo>.
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The sRNA is coupled to a strong promotor, J23101
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===Reference===
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Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli
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Vandana Sharma, Asami Yamamura, and Yohei Yokobayashi
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ACS Synthetic Biology 2012 1 (1), 6-13
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<partinfo>BBa_K864440 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K864440 SequenceAndFeatures</partinfo>
  
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===References===
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[http://www.ncbi.nlm.nih.gov/pubmed/12101127] Møller, T., Franch, T., Udesen, C., Gerdes, K., and Valentin-Hansen, P. (2002) Spot 42 RNA mediates discoordinate expression of the E. coli galactose operon Genes Dev. 16, 1696– 1706
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[http://pubs.acs.org/doi/full/10.1021/sb200001q] Sharma V., Yamamura A., and Yokobayashi, Y. (2012): Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli. ACS Synthetic Biology 1 (1), 6-13
  
 
<!-- Uncomment this to enable Functional Parameter display  
 
<!-- Uncomment this to enable Functional Parameter display  

Latest revision as of 18:15, 29 September 2012

Small RNA spot42

This part is a component of the Uppsala University iGEM 2012 modular small RNA screening system, together with the target construct BBa_K864444.

This is the small RNA spot42 from E coli MG1655 coupled to the strong promoter BBa_J23101. The spot42 regulates the galactose metabolism by binding to the mRNA and recruiting Hfq proteins [1], blocking initiation of translation. The spot42 has proven to be an especially suitable sRNA for engineering of silencing sRNA:s, by replacing its RNA-binding domain[2].

To create a random sRNA library, the RNA-binding domain can be randomized by site-directed mutagenesis using the primers BBa_K864445 and BBa_K864446.


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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

[http://www.ncbi.nlm.nih.gov/pubmed/12101127] Møller, T., Franch, T., Udesen, C., Gerdes, K., and Valentin-Hansen, P. (2002) Spot 42 RNA mediates discoordinate expression of the E. coli galactose operon Genes Dev. 16, 1696– 1706

[http://pubs.acs.org/doi/full/10.1021/sb200001q] Sharma V., Yamamura A., and Yokobayashi, Y. (2012): Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli. ACS Synthetic Biology 1 (1), 6-13