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 metabolic pathways. The spot42 scaffold recruits Hfq proteins. In some cases it has been shown to bind to the ribosomal binding site of the mRNA and thereby blocking translation.  ( 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 down regulation 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 that could inhibit translation of a gene in question.
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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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===References===
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Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli
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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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Vandana Sharma, Asami Yamamura, and Yohei Yokobayashi
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ACS Synthetic Biology 2012 1 (1), 6-13
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[http://pubs.acs.org/doi/full/10.1021/sb200001q] Sharma V., Yamamura A., and Yokobayashi, Y. (2002): Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli. ACS Synthetic Biology 2012 1 (1), 6-13
  
  

Revision as of 16:21, 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.

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. (2002): Engineering Artificial Small RNAs for Conditional Gene Silencing in Escherichia coli. ACS Synthetic Biology 2012 1 (1), 6-13


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]