Part:BBa_J364000

Test Device 1 for the iGEM InterLab Study

This is a GFP expressing constitutive device for the 2017 iGEM InterLab study. It is called Test Device 1 for the study for easy reference.

This device is stored in pSB1C3 for the InterLab and is fully BioBrick compatible.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 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 705
  

Team NAWI_Graz 2019: GFP dynamics of transformed Escherichia coli strains in LB broth

Background

In Team NAWI_Graz 2019 characterization, we found that in normal growth/incubation condition (37 ^oC, LB agar) BBa_J364000-transformed Escherichia coli BL21 cells had the highest GFP expression in comparison to BL21 Star and Top 10 strains. In order to confirm that we have measured the GFP Flourescence of those three strains over 44 hours period.

Experimental Design

We used three different strains of transformed E. coli (Top10,BL21 and BL21 Star) for this study. Transformed strains were incubated in 15 mL LB broth 37 ^oC overnight. Next day they were used to inoculate a 50 mL LB to OD₆₀₀=0.1. The cultures were then incubated at 37 ^oC, 140 rpm and samples were taken every 4 hours for 2 days to determine the green fluorescence protein (GFP) fluorescence intensity at 510 nm.

Result and Findings

• There are significant differences of GFP expression in different strains of E. coli (Top10, BL21, BL21 Star)
• The broth became ligh green in color under natural light after around 14-18 hours of incubation time.

          Figure 1: GFL Flourescence and OD600 Measurements with GFP containing E.coli strains and non GFP containing E.coli strains (control))

Figure 2: ~14 hours old GFP-transformed E. coli strains (Left: Top10, Middle: BL21, Right: BL21 Star)