Part:BBa_K2876005
A fusion of a RNAPolIII alpha subunit and antiAFB1-ScFv1
A fusion of a RNAPolIII alpha subunit and an aflatoxin sensing single chain antibody variable region (antiAFB1-ScFv1). Used in conjunction with BBa_K2876003 (https://parts.igem.org/Part:BBa_K2876005) and BBa_K2876000 (https://parts.igem.org/Part:BBa_K2876000) to create for an E. coli two hybrid system capable of detecting aflatoxin.
For our small molecule test case we chose to detect aflatoxin, a small molecule produced by Aspergillus fungi species that infects many agricultural crops, including cannabis.2,3 Aflatoxin also causes a number of health problems in people when inhaled or ingested by immunocompromised patients.7 We fused two single chain antibodies targeting aflatoxin (BBa_K2876005 and BBa_K2876003) to the lambda repressor and alpha subunit of RNA-polymerase III (BBa_K2876002) described in Dove, Joung, and Hochschild in order to create two fusion proteins capable of activating transcription in the presence of aflatoxin (Figure 4 small molecule). For our reporter we used the previously described pOL2-62 from Dove, Joung, and Hochschild fused to a RFP. See more information about our reporter below.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 8
Illegal NotI site found at 766 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 480
- 1000COMPATIBLE WITH RFC[1000]
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