Difference between revisions of "Part:BBa K1899007"

Latest revision as of 15:46, 18 October 2016

phlFp-B0032-Lacl-B0032-TetR-GFP

This construct is designed to estimate the workload of E. coli and measure the fluorescence expression levels by the promoter when it is ligated to the tristable switch.

Results

The fold change between construct A (pSB3K3-BBa_phlFp-E0240) and negative control (pSB3K3-BBa_E0240), and that of construct E (pSB3K3-BBa_phlFp-B0032-C0012-B0032-C0040-E0240) is 13.2 and 1.8 times respectively.

Owing to the increase in number of coding regions, the workload of the cells is highly increased. The production rate of fluorescence should therefore be lower. It is acceptable to obtain a drop in the RFU level between the two constructs.

Fig 1. Comparison on fluorescence expression levels of construct A (BBa_phlFp-E0240) and construct E (BBa_phlFp-B0032-C0012-B0032-C0040-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
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1240
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 2629

@@ Line 7: / Line 7: @@
 ===Results===
+The fold change between construct A (pSB3K3-BBa_<i>phlFp</i>-E0240) and  negative control (pSB3K3-BBa_E0240), and that of construct E (pSB3K3-BBa_<i>phlFp</i>-B0032-C0012-B0032-C0040-E0240) is 13.2 and 1.8 times respectively.
-https://static.igem.org/mediawiki/parts/thumb/9/95/Comparison_of_Construct_A_and_E.jpeg/693px-Comparison_of_Construct_A_and_E.jpeg
+<br><br>
+Owing to the increase in number of coding regions, the workload of the cells is highly increased. The production rate of fluorescence should therefore be lower. It is acceptable to obtain a drop in the RFU level between the two constructs.
-Fig a ) <b>Comparison of Technical Triplicate Results of pSB3K3-<i>phlFp</i> - BBa_B0032-C0012-B0032-C0040-E0240(E) and pSB3K3-<i>phlFp</i>- BBa_E0240(A)</b>
+<br>
+[[File:IGEM2016 HKUST phlFIK1899007.png|thumb|600px|center|<b>Fig 1. Comparison on fluorescence expression levels of construct A (BBa_<i>phlFp</i>-E0240) and construct E (BBa_<i>phlFp</i>-B0032-C0012-B0032-C0040-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.]]
-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-<i>phlFp</i>-BBa_B0032-C0012-B0032-C0040-E0240(E)</b> is 13.2 and 1.8 times respectively.
-Owing to the increase of the number of coding regions, the workload of the cells is highly increased. The production rate of fluorescence should therefore be lower. It is acceptable to obtain a drop in the RFU level of<b> pSB3K3-<i>phlFp</i>-BBa_B0032-C0012-B0032-C0040-E0240(E)</b> comparing with <b>pSB3K3-<i>phlFp</i>- BBa_E0240(A)</b>.