Part:BBa_K4634015
T7ptag
We constructed an AND gate in E. coli that takes two stimulus signals from the environment as input, and the two signals activate the transcription of their respective regulatory genes. The AND gate can only be triggered when both regulatory genes are transcriptionally active and integrated with each other. Integration occurs through interactions between mRNA and tRNA. The first regulatory gene is T7ptag (T7 RNA polymerase TAG); the second regulatory gene is tRNA supD, which is specifically used for the translation of T7ptag. When both regulatory genes are transcribed, T7ptag is translated and synthesized. T7ptag can specifically bind to the T7 promoter and translate the genes downstream of the T7 promoter (i.e., the genes regulated by the AND gate), thereby realizing the AND gate. Function[1]. As one of the elements that implements the AND gate, T7ptag is different from the general T7 RNA polymerase in that the former has been modified to contain two amber stop codons that prevent translation (replaced by TAG at the original 13 and 204 amino acid positions). These stop codons are translated as serine when supD is also transcribed, continuing the translation process that should have been terminated. As a result: when T7ptag and supD are transcribed at the same time, T7ptag can be translated and synthesized, and the genes downstream of the T7 promoter can be expressed (Fig. 1) [1].
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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