Difference between revisions of "Part:BBa K1899005"

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:

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 1539

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 <br>
 ===Results===
-The fold change between<b> pSB3K3-<i>phlFp</i> - BBa_E0240(A)</b>, negative control<b> (pSB3K3-BBa_E0240)</b> and that of <b>pSB3K3-BBa_J23101-B0032-C0040-B1006- <i>phlFp</i> -E0240(D)</b> is around 13.2 times and 7.01 times respectively.
+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>
-This is 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 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.
+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.
 [[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.]]