Part:BBa_K1439001

This part contains a reporter gene BBa_J04450, combined with OriTRP4. Used to test plasmid mobility.

Once the conjugation take place,the recipient bacterium will obtain the reporter gene which gives us a signal to distinguish whether the conjugation has happened as well as the conjugation efficiency easily through observation. In addition, it facilitates the Follow-up experiments on the conjugation rate, conjugation time, and conjugation efficiency between different strains.

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 NgoMIV site found at 20
  Illegal NgoMIV site found at 30
  Illegal AgeI site found at 1139
  Illegal AgeI site found at 1251
  
• 1000
  COMPATIBLE WITH RFC[1000]

Result

We do conjugation experiment between HB101 and Top10 to test the conjugation ability of OriTRP4 (BBa_K1439000). The HB101 strain, sensitive to streptomycin, is provided by QIU Zhigang. And Top10 has streptomycin resistance. In order to test BBa_K1439000 better, we ligate OriTRP4 with the report gene RFP. After conjugation, we pick the red colonies by LB medium with resistance.

Conjugate between E.coil HB101 and E.coil Top10.