Part:BBa_K2708003

Promoter and S-adenosyl methionine riboswitch, GFP fluorescent expression system.

Characterizing Part:

Here's a map of its gene sequencing,99% are consistent.

The Genes line consists of the promoter, the S-adenosyl methionine riboswitch and promoter E0840.The plasmid is converted into E.coli to measure green fluorescence to verify the riboswitch's function.When the circuit is activated by the promoter, the terminating substructure at the end of the SAM ribose switch cannot be formed, making the circuit express downward to produce GFP fluorescent protein.While the molecule SAM binds to the SAM riboswitch, the end of the SAM riboswitch forms a terminating substructure that terminates the circuit and fails to express the fluorescent protein.

Its size was 1176bp, and after adding the enzyme cutting sites at both ends, it was between 1500bp and 2000bp. Agarose gel electrophoresis showed the bands correctly.

Experiment Results :

1.In the higher SAM concentration range: The experimental results showed that the higher the concentration of SAM, the lower the fluorescence and OD600 value, indicating the response of the SAM switch to SAM, which led to the inhibition of fluorescence expression behind the circuit ,as expected.

2.In the lower SAM concentration range: The experimental results showed that the higher the concentration of SAM, the lower the fluorescence and OD600 value, indicating the response of the SAM switch to SAM, which led to the inhibition of fluorescence expression behind the circuit ,as expected.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 963