Difference between revisions of "Part:BBa K1899008"

Latest revision as of 16:05, 18 October 2016

phlFp-E0240

Construct for characterization

Fig.1 phlFp with reporter gene.

In order to compare the promoter strength of the phlFp with the other two promoters, tetp and lacp, it was ligated with a GFP reporter gene, BBa_E0240, in BioBrick RFC10 standard. With the presence of a green fluorescence protein reporter gene, GFP protein would be produced during the characterization, thus, the strength of phlFp could be compared to that of the other two promoters in our project, lacp and tetp.

Results

Experiments on comparing the strength of phlFp, tetp and lacp, were performed. Results indicated that phlFp is the strongest among the three, with 1.96 times and 6 times stronger than that of tetp and lacp, respectively.

Fig 2. Comparison on the strength of phlFp, tetp and lacp. 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
COMPATIBLE WITH RFC[12]
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 737

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1899008 short</partinfo>
-With all the essential elements for a circuit to function, this part could be used independently in the project to estimate the promoter stength of the phlFp. Since it contains a green fluorescence protein reporter gene, GFP protein would be produced during the characterisation, thus, the intensity of the GFP could be used to estimate the phlFp promoter strength and compare it to that of the other two promoters in our project, lacp and tetp.
+==Construct for characterization==
+[[File:PhlFp-E0240_2.png|thumb|500px|center|<b>Fig.1 </b> <i>phlFp</i> with reporter gene.]]
+In order to compare the promoter strength of the <i>phlFp</i> with the other two promoters, <i>tetp</i> and <i>lacp</i>, it was ligated with a GFP reporter gene, BBa_E0240, in BioBrick RFC10 standard. With the presence of a green fluorescence protein reporter gene, GFP protein would be produced during the characterization, thus, the strength of <i>phlFp</i> could be compared to that of the other two promoters in our project, <i>lacp</i> and <i>tetp</i>.
+===Results===
+Experiments on comparing the strength of <i>phlFp</i>, <i>tetp</i> and <i>lacp</i>, were performed. Results indicated that <i>phlFp</i> is the strongest among the three, with 1.96 times and 6 times stronger than that of <i>tetp</i> and <i>lacp</i>, respectively.
+[[File:IGEM2016 HKUST constructA.png|thumb|600px|center|<b>Fig 2. Comparison on the strength of <i>phlFp</i>, <i>tetp</i> and <i>lacp</i>.</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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