Difference between revisions of "Part:BBa J364000"
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<partinfo>BBa_J364000 parameters</partinfo>
 
<partinfo>BBa_J364000 parameters</partinfo>
 
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==Team NAWI_Graz 2019: GFP dynamics of cloned <i>Escherichia coli</i> strains in LB broth==
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<html>
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<p>
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<figure>
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    <img src="https://static.igem.org/mediawiki/parts/a/a5/GFP_Absorbance_510.png" alt="Three strains expression on GFP and its control" style="float:right;width:350px;height:205.5px;">
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</figure>
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<i>Background</i><br><br>
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In <a href="https://2019.igem.org/Team:NAWI_Graz" target="_blank">Team NAWI_Graz 2019</a>
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characterization, we found in normal growth/incubation condition (37 <sup>o</sup>C, LB agar) that
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BBa_J364000-transformed <i>Escherichia coli</i> BL21 colony 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. <br><br>
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<i>Experimental Design</i><br><br>
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We used three different strains of transformed <i>E. coli</i> (Top10,BL21 and BL21 Star) for this study.
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Transformed strain were incubated in 15 mL LB broth 37 <sup>o</sup>C overnight. Next day they were used to inoculate a 50 mL LB to OD<sub>600</sub>=0.1. The cultures were then incubated at 37 <sup>o</sup>C, 140 rpm and were sampled every 4 hours for 2 days to determine the green fluorescence protein (GFP) absorbance at 510 nm.
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<br><br>
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<i>Result and Findings</i><br>
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<ul>
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  <li>There are significant differences of GFP expression in different strains of <i>E. coli</i>
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(Top10, BL21, BL21 Star)</li>
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  <li>The broth become ligh green in color under natural light around 14-18 hours of incubation time.</li>
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</ul>
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<br>
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<img src="https://static.igem.org/mediawiki/parts/8/82/Fluorescencetop10%2Bk.png" style="width: 310px; height: 205.5;">
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<img src="https://static.igem.org/mediawiki/parts/2/2b/Bl21star_fluorescence_%2Bcontrol.png" style="width: 310px; height: 205.5;">
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<img src="https://static.igem.org/mediawiki/parts/e/e7/Gfp_bl21_fluoreseecnece.png" style="width: 310px; height: 205.5;">
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<br>
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<img src="https://static.igem.org/mediawiki/parts/e/ea/Top10%2Bcontrol.png" style="width: 310px; height: 205.5;">
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<img src="https://static.igem.org/mediawiki/parts/0/02/Bl21star%2Bcontrol.png" style="width: 310px; height: 205.5;">
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<img src="https://static.igem.org/mediawiki/parts/3/32/Bl21%2Bcontrol.png" style="width: 310px; height: 205.5;">
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<figcaption> <i><b>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;Figure 1:</b> GFL Flourescence and OD600 Measurements with GFP containing E.coli strains and non GFP containing E.coli strains (control))</i></figcaption></figure>
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<br> <br>
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<figure><img src="
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                    https://static.igem.org/mediawiki/parts/3/3b/14h_flask_photo.jpeg" style="height: 30%; width: 30%;"/>
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                                          <figcaption><i><b>Figure 2:</b> ~14 hours old GFP-transformed E. coli strains (Left: Top10, Middle: BL21, Right: BL21 Star)</i></figcaption></figure>
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</html>

Revision as of 10:37, 15 October 2019


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 cloned Escherichia coli strains in LB broth

Three strains expression on GFP and its control
Background

In Team NAWI_Graz 2019 characterization, we found in normal growth/incubation condition (37 oC, LB agar) that BBa_J364000-transformed Escherichia coli BL21 colony 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 strain 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 were sampled every 4 hours for 2 days to determine the green fluorescence protein (GFP) absorbance 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 become ligh green in color under natural light 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)