Part:BBa_K5097002
pH Response Element riboswitch m13
In 1990, Bingham et al. identified a locus within E.coli whose activity was tied to pH changes, which they coined Alx. It exhibited increased expression at pH 8.5 (1). This activity was further characterized by Nechooshtan et. al., who determined that the Alx promoter and riboswitch are responsible for increased Alx expression in response to high pH (2). The Alx promoter and riboswitch have been previously entered into the parts repository by Team NAXI_GRAS under part number BBa_K2348000.
Our team redesigned the pH-responsive element (PRE) (Bba_K5097000) from the Alex locus by isolating the riboswitch sequence to allow regulation of the riboswitch independent of the promoter. We utilized the PRE sequence published by (3) to delineate the riboswitch sequence. And cloned it into a reporter gene construct (Bba_ K5097005) with BFP to test the pH responsiveness and its control of translation of the BFP gene.
After screening clones via colony PCR we sent clones for Sanger sequencing to confirm the sequence of the riboswitch. We identified a number of mutations in the clones, including one, that we designated m13. We performed a pairwise alignment to compare the sequence of m13 to PRE, and identified three deletions within the sequence.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 116
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