Part:BBa_K2392000
Aflatoxin B1 Biosensor Expressor Component
This part is the expressor component of the UGA iGEM Team's Aflatoxin B1 Biosensor. The two-component biosensor is based on of a riboswitch design where this repressor acts bind to the expressor component, which fluoresces in the presence of Aflatoxin B1. This expressor is composed of a promoter, DNA complementary (cDNA) to the aflatoxin B1 aptamer, a ribosome binding site (RBS), green fluorescent protein (GFP), and a terminator. This part works by fluorescing in the presence of Aflatoxin B1, while normally being repressed by the repressor under conditions without Aflatoxin B1. In the presence of Aflatoxin B1, the repressor part will theoretically bind preferentially to Aflatoxin B1 rather than to the expressor. This part has been tested to be specifically repressed by our repressor part when transformed together in vivo. This part works as an aptamer-based expressor for an Aflatoxin B1 biosensor. This working expressor is a modified version of an original design that was non-functional. The original design's lack of fluorescence was likely due to the occlusion of the RBS in a stem-loop structure as seen in Figure 1. Truncating the cDNA region by 9 bases from the 3' end disrupted this secondary structure (as seen in Figure 2.) and resulted in this functional expressor.
Figure 1. Secondary structure of the cDNA and RBS region of the original expressor design.
Figure 2. Secondary structure of the cDNA and RBS region of a modified expressor, E3.
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 731
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