Difference between revisions of "Part:BBa K1899005"
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<partinfo>BBa_K1899005 short</partinfo> | <partinfo>BBa_K1899005 short</partinfo> | ||
| − | This part is constructed for investigating the effect of | + | This part is constructed for investigating the effect of TetR over <i>phlFp</i>. |
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===Results=== | ===Results=== | ||
| − | + | The fold change between construct A (pSB3K3-BBa_<i>phlFp</i> -E0240) and negative control (pSB3K3-BBa_E0240), and that of construct D (pSB3K3-BBa_J23101-B0032-C0040-B1006- <i>phlFp</i>-E0240) is around 13.2 times and 7.01 times respectively. | |
| + | <br><br> | ||
| + | We hypothesized that this may due to the toxicity of <bbpart>BBa_C0040 </bbpart> (TetR). The toxicity reduces the growth rate of the <i> E. coli</i> containing this plasmid. The strong promoter <bbpart>BBa_J23101</bbpart> makes this effect more significant when doing the characterisation. Though all the data were collected with OD<sub>600</sub> within the mid-log range, there is still a variation between them, making a significant difference in the RFU. It is ungrounded to say TetR interferes the functionality of <i> phlFp</i> at this stage. | ||
| − | <b>Fig | + | [[File:IGEM2016 HKUST pphlFK1899005.png|thumb|600px|center|<b>Fig 1. Comparison on fluorescence expression levels of construct A (BBa_<i>phlFp</i>-E0240) and construct D (BBa_J23101-B0032-C0040-B1006- <i>phlFp</i> -E0240)</b>. Negative control represents BBa_E0240. Characterization was done using <i>E. coli</i> strain JW0336. Cells were first precultured overnight and were subcultured to mid-log phase where GFP emission measurements were made using an EnVision® multilabel reader. This result was obtained by combining 3 characterization data obtained in 3 different days. Error bar present SD from 3 biological replicates.]] |
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<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
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<partinfo>BBa_K1899005 SequenceAndFeatures</partinfo> | <partinfo>BBa_K1899005 SequenceAndFeatures</partinfo> | ||
Latest revision as of 15:17, 18 October 2016
J23101-B0032-TetR-B1006- phlFp -GFP
This part is constructed for investigating the effect of TetR over phlFp.
Results
The fold change between construct A (pSB3K3-BBa_phlFp -E0240) and negative control (pSB3K3-BBa_E0240), and that of construct D (pSB3K3-BBa_J23101-B0032-C0040-B1006- phlFp-E0240) is around 13.2 times and 7.01 times respectively.
We hypothesized that this may due to the toxicity of BBa_C0040 (TetR). The toxicity reduces the growth rate of the E. coli containing this plasmid. The strong promoter BBa_J23101 makes this effect more significant when doing the characterisation. Though all the data were collected with OD600 within the mid-log range, there is still a variation between them, making a significant difference in the RFU. It is ungrounded to say TetR interferes the functionality of phlFp at this stage.
Fig 1. Comparison on fluorescence expression levels of construct A (BBa_phlFp-E0240) and construct D (BBa_J23101-B0032-C0040-B1006- phlFp -E0240). Negative control represents BBa_E0240. Characterization was done using E. coli strain JW0336. Cells were first precultured overnight and were subcultured to mid-log phase where GFP emission measurements were made using an EnVision® multilabel reader. This result was obtained by combining 3 characterization data obtained in 3 different days. Error bar present SD from 3 biological replicates.
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 1539