Part:BBa_K2595004
Spot 42
Encoded by the spf gene in 5 ɣ-proteobacterial orders, Spot 42 is an 109 nucleotide long antisense sRNA that can effectively downregulate the expression of gal operon.(P. Haugen et. al, 2015) Spot 42 sRNA is composed of 3 regions: an unstructured loop for target binding, as well as a Rho-independent terminator and AU-rich terminator that contribute to structural stability of the polynucleotide. Spot 42 exerts its impact by binding to post-transcriptional regulator Hfq which in turn facilitates the hybridization of Spot 42 to Shine-Dalgarno region of galK, resulting in a decrease in galactokinase production. The expression of Spot 42 is highly dependent on intracellular glucose and cAMP level as cAMP-CRP complex is a main suppressor of spf expression. (GA Hansen et al., 2012) Studies also reveal that Spot 42 is likely to interact with mK2 mRNA and affect galT expression, which could add to galK inhibition. (Wang et al., 2015)
Usage and Biology
This synthetic version of Spot 42 follow the same principles as its wild type partner to regulate gene expression. This modified construct, however, can target any gene of interest due to its sequence which contains a BaeI restriction enzyme site. After digestion with this restriction enzyme and ligation with a target sequence, Spot 42 can regulate the expression of the gene that contains the complementary regions for its targeting sequence. Ligation with the desired targeting sequence does not create any scar in the sRNA sequence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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