Difference between revisions of "Part:BBa K1899009"

Latest revision as of 21:28, 19 October 2016

B0032-LacI

LacI is conjugated to a medium strength RBS

Construct for Characterisation

Fig.1 lacI-lacp with reporter gene.

In order to characterise the efficiency of this construction intermediate, it was ligated with a construct containing lacp and GFP reporter gene, BBa_E0240. The expression of LacI was driven by a strong constitutive promoter, BBa_J23101, and terminated by BBa_B1006 terminator. The assembly processes were performed in BioBrick RFC10 standard.

Results

Experiments on comparing the GFP expression driven by lacp from constructs with and without BBa_B0032-lacI were performed. Negative control used was BBa_E0240. Results indicated that BBa_B0032-lacI driven by BBa_J23101 and terminated by BBa_B1006 could reduce the GFP expression by 2.9 times.

Fig 2. Comparison on the GFP expression of construct with and without BBa_B0032-lacI. BBa_E0240 was used as negative control. Characterisation was done using E. coli strain JW0336. Cells were first pre-cultured 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 characterisation 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
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1166
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 6: / Line 6: @@
 ==Construct for Characterisation==
-[[File:BBa_K1899009_lacI-lacp-E0240.png|thumb|800px|center|<b>Fig.1 </b> <i>tetR</i>-<i>tetp</i> with reporter gene.]]
+[[File:BBa_K1899009_lacI-lacp-E0240.png|thumb|800px|center|<b>Fig.1 </b> <i>lacI</i>-<i>lacp</i> with reporter gene.]]
-In order to characterise the efficiency of this construction intermediate, it was ligated with a construct containing <i>lacp</i> and GFP reporter gene, BBa_E0240. The expression of LacI was driven by a strong constitutive promoter, BBa_J23101, and terminated by BBa_B1006 terminator. The assembly processes are performed in BioBrick RFC10 standard.
+In order to characterise the efficiency of this construction intermediate, it was ligated with a construct containing <i>lacp</i> and GFP reporter gene, BBa_E0240. The expression of LacI was driven by a strong constitutive promoter, BBa_J23101, and terminated by BBa_B1006 terminator. The assembly processes were performed in BioBrick RFC10 standard.
 ===Results===
-Experiments on comparing the GFP expression driven by <i>lacp</i> from constructs with and without BBa_B0032-<i>tetR</i> were performed. Negative control used was BBa_E0240. Results indicated that BBa_B0032-<i>lacI</i> driven by BBa_J23101 and terminated by BBa_B1006 could reduce the GFP expression by xx times.
+Experiments on comparing the GFP expression driven by <i>lacp</i> from constructs with and without BBa_B0032-<i>lacI</i> were performed. Negative control used was BBa_E0240. Results indicated that BBa_B0032-<i>lacI</i> driven by BBa_J23101 and terminated by BBa_B1006 could reduce the GFP expression by 2.9 times.
-[[File:BBa K1899010-iGEM16 Hong Kong HKUST Characterisation.jpeg|thumb|600px|center|<b>Fig 2. Comparison on the GFP expression of construct with and without BBa_B0032-<i>tetR</i>.</b> BBa_E0240 was used as negative control. 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:BBa K1899009-iGEM16 Hong Kong HKUST Characterisation.jpeg|thumb|600px|center|<b>Fig 2. Comparison on the GFP expression of construct with and without BBa_B0032-<i>lacI</i>.</b> BBa_E0240 was used as negative control. Characterisation was done using <i>E. coli</i> strain JW0336. Cells were first pre-cultured 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 characterisation data obtained in 3 different days. Error bar present SD from 3 biological replicates.]]