Part:BBa_K864440

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]