Part:BBa_K4438601

T6_Trigger_1_midtrigger_phi29

T6_Trigger_1_midtrigger_phi29 is single-stranded DNA having 27 nucleotides. Figure 1B) Illustrates the secondary structure and its minimum free energy.

Usage and Biology

Few bases of this part are complementary (Watson-Crick base pairing) to T6 BBa_K4438600 and are designed to block its sequence motif. Motif, mostly associated with the loops, is linked to the formation of the specific binding site for testosterone [1]. Figure 1D) Shows the secondary structure of both parts hybridised at 37° Celsius. The hormone binds with T6 BBa_K4438600 with high affinity and displaces the T6_Trigger_1_midtrigger_phi29. This part has complete complementarity with the part T6_Binder_1(BBa_K4438602). Phi 29 DNA extension polymerase extends the template strand and in-vitro transcription of the duplex forms multiple broccoli light-up aptamers. Different levels of testosterone, an anabolic androgenic steroid (AAS), in biological fluids can be detected using all these three parts.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

Skouridou, V., Jauset-Rubio, M., Ballester, P., Bashammakh, A. S., El-Shahawi, M. S., Alyoubi, A. O., & O’Sullivan, C. K. (2017). Selection and characterization of DNA aptamers against the steroid testosterone. Microchimica Acta, 184(6), 1631-1639.