Part:BBa_K117004

pLacI-GFP

The construct design

‎BBa K117004

This biobrick device has a lacI regulated promoter which upon the induction of lactose/IPTG, will casue the expression of the reporter gene, Green fluorescence protein (GFP EE0040).

Abnormality found in the registry

This device could be found in the registry in the form of BBa_J04430. However, the team found an abnormality when BBa_J04430 was transformed into Top 10 cells. As shown on the pictures below, the cells glow pink instead of green under UV light.

BBa_J04430 glows pink

Making our own part

Hence the team constructed this part from the registry by ligating BBa_R0010 with BBa_E02040. The BBa_E02040 consist of the biobrick parts: BB0034, E0040, B0010 and B0012.

Characterization of pLacI-GFP

The NTU@iGEM team characterized the biobrick part pLacI-GFP. The influence of 1)concentration of Lactose 2) temperature and 3) time were investigated.

Shown below is the graph obtained for the experiment conducted at 37°C.

Characterization at 37°C

Electron microscope pictures

Cells induced with 1mM lactose concentration

Cells induced with 10mM lactose concentration

For more information including the protocols and results obtained, please go to: [http://2008.igem.org/Team:NTU-Singapore/Parts/Characterization_of_LacI-GFP NTU@iGEM Characterization of pLac-GFP]

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 873