Part:BBa_K1714001

pLac-RBS B0034-ABF-2-double terminator

This part will be finally introduced upstream of BBa_K1714004 (PBAD-B0034-BLA-dT on 1C3) and expressed as AMPs in Co-expression system. In this page, I described the result of the activity assay of ABF-2.

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
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 321
  
• 1000
  COMPATIBLE WITH RFC[1000]

In this page, I described the result of the activity assay of ABF-2.

We transformed it into E. coli (in this case, we used DH5α) and compared the growth curve between samples induced or not. Activity test for ABF-2 and thanatin →Comparing growth curve between the samples one is induced by IPTG and the other is not induced.

1. Diluting the bacterial culture which is enough pre-cultured by 0.1 OD₆₀₀ (Optical Density in λ=600nm) in 2 mL LBC.(Lysogeny Broth liquid culture including chloramphenicol)
2. Adding 2 µL of IPTG into one and DW into the other.
3. Measuring OD₆₀₀ of these samples once in an hour.

As a result, we got the graph of growth curve of DH5α and shown on below as Chart1.

Fig. 8. The growth curve of E. coli including BBa_K1714001 with IPTG or not.

As you can see, we can’t say that ABF-2 in BBa_K1714001 has correct antimicrobial activity.