Part:BBa_K4438603

T6_Trigger_2_phi29

T6_Trigger_2_phi29 is single-stranded DNA having 30 nucleotides. The 5’ end is flanked by a few bases that are complementary to the sense T7 promoter sequence. 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 2D) 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_2_phi29. This part has complete complementarity with the part T6_Binder_2(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.