Part:BBa_K5299118
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T7 artificial terminator (T7hyb6)
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Secondary Structure
Discreption
The T7hyb6 terminator was designed by combining hairpin-forming T7RNAP terminators and a sequence-dependent pause site (from the vesicular stomatitis virus (VSV sequence): TATCTGTT). The VSV sequence was inserted in the poly-U-proximal segment of the terminator stem as 2 copies that overlap by 1 base pair. Calvopina-Chavez et al. showed with in vivo testing that it has a termination efficiency of 91% compared to the native terminator that had 62%. They also tested the termination efficiency for E. coli RNAP and it showed an efficiency of 47% compared to the native that had 83% [1].
Use
The native T7 bacteriophage terminator allows read-through transcription causing the transcripts to be much longer, so terminators with better termination efficiency are needed. This engineered terminator outperforms the efficiency of the native terminator. Because of that we deemed it suitable to use it in our project for the production of dsRNA molecules of specific length in P. putida.
Measurement
References
[1] Calvopina-Chavez, D. G., Gardner, M. A., & Griffitts, J. S. (2022). Engineering efficient termination of bacteriophage T7 RNA polymerase transcription. G3 (Bethesda, Md.), 12(6). doi:10.1093/g3journal/jkac070
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