Difference between revisions of "Part:BBa K1899005"
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===Results===
 
===Results===
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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.
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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.
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[[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.]]
 
[[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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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. Mean and standard error mean (SEM) are calculated and represented by the height of the bar and the error bar respectively.
 
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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.
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 15:04, 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 pSB3K3-phlFp - BBa_E0240(A), negative control (pSB3K3-BBa_E0240) and that of pSB3K3-BBa_J23101-B0032-C0040-B1006- phlFp -E0240(D) is around 13.2 times and 7.01 times respectively.

This is 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 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 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