Part:BBa_K4813009
HxlR K13A - mutated HxlR codon with enhanced DNA binding and transcription activation
The coding sequence mentioned encodes a protein called HxlR, which is a formaldehyde-responsive transcription factor from Bacillus subtilis. According to research, when the 13th amino acid in HxlR protein, Lysine, was replaced by Alanine, mutated HxlR-K13A showed a more optimal conformation for DNA binding activity and a higher transcription activation than the wild-type HxlR protein [1].
In our project, we incorporated the mutated K13A version of HxlR gene into two composite parts to examine if HxlR-K13A can lead to an enhanced expression of our reporter gene, a red-colour chromoprotein called dTomato under the presence of formaldehyde. In one composite part, HxlR-K13A expression is driven by a strong and constitutive reverse promoter (BBa_J23100) for HxlR-K13A to be backward transcribed like the wild-type HxlR. As for the other composite part, HxlR-K13A expression is driven by a T7 promoter to be forward transcribed.
References
[1]Zhu, Rongfeng, et al. “Genetically encoded formaldehyde sensors inspired by a protein intra-helical crosslinking reaction.” Nature Communications, vol. 12, no. 1, 2021, https://doi.org/10.1038/s41467-020-20754-4
Usage and Biology
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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