Difference between revisions of "Part:BBa K1033225:Experience"
(→User Reviews) |
(→Applications of BBa_K1033225) |
||
| Line 4: | Line 4: | ||
===Applications of BBa_K1033225=== | ===Applications of BBa_K1033225=== | ||
| + | <html> | ||
| + | |||
| + | <I>BABS UNSW 2015</I> | ||
| + | <br>A LacZ assay was carried out with p170 (<a href="https://parts.igem.org/Part:BBa_K1677300">BBa_K1677300</a>), CP44 (<a href="https://parts.igem.org/Part:BBa_K1677300">BBa_K1677300</a>) and ASR (<a href="https://parts.igem.org/Part:BBa_K1033225">BBa_K1033225</a>) fused to a promoter activity reporter (BBa_I732095) in E. Coli and grown in LB with antibiotics for 8 hours. The data normalized to OD600 of the cultures reveals a decrease in lacZ production correlated with a decrease in pH for the CP44 promoter. A slight increase was observed in both p170 and ASR with the decline in pH, however expression of lacZ is much lower even at neutral pH than with CP44. </br> | ||
| + | |||
| + | <figure><center><img src="https://static.igem.org/mediawiki/2015/a/af/Unsw2015_laczassay_ph_odnormalised_graph.jpeg" style="PADDING-BOTTOM:0%;PADDING-TOP:0%; max-width:70%;"></center></figure> | ||
| + | |||
| + | |||
| + | <br> When the above data is normalized to the neutral pH value of the culture the differential expression at varying pH values becomes more pronounced. Both the E. Coli and L. Lactis ASR promoters show an increase in activity at lower pH values. <br> While ASR was proven to have increased levels of RNA at lowered pH, the biobrick had not previously had the activity of the promoter characterized. | ||
| + | |||
| + | <figure><center><img src="https://static.igem.org/mediawiki/2015/a/ac/Unsw2015_laczassay_ph_column_graph.jpeg" style="PADDING-BOTTOM:0%;PADDING-TOP:0%; max-width:70%;"></center></figure> | ||
| + | |||
| + | </html> | ||
===User Reviews=== | ===User Reviews=== | ||
Revision as of 18:35, 18 September 2015
This experience page is provided so that any user may enter their experience using this part.
Please enter
how you used this part and how it worked out.
Applications of BBa_K1033225
BABS UNSW 2015
A LacZ assay was carried out with p170 (BBa_K1677300), CP44 (BBa_K1677300) and ASR (BBa_K1033225) fused to a promoter activity reporter (BBa_I732095) in E. Coli and grown in LB with antibiotics for 8 hours. The data normalized to OD600 of the cultures reveals a decrease in lacZ production correlated with a decrease in pH for the CP44 promoter. A slight increase was observed in both p170 and ASR with the decline in pH, however expression of lacZ is much lower even at neutral pH than with CP44.
When the above data is normalized to the neutral pH value of the culture the differential expression at varying pH values becomes more pronounced. Both the E. Coli and L. Lactis ASR promoters show an increase in activity at lower pH values.
While ASR was proven to have increased levels of RNA at lowered pH, the biobrick had not previously had the activity of the promoter characterized.
User Reviews
UNIQ61d7e207b5d32d50-partinfo-00000001-QINU
|
No review score entered. Username |
This part was used in the composite part BBa_K1365555 for characterization by the Rijksuniversiteit Groningen.
The graph shown above helps us to know distribution of the fluorescence in the cells of E.coli with different CP promotor fused to superfoldedGFP-terminator-RBS collection (CP1, CP11, CP29, CP30, CP41, CP44) and J23101 as reference. We used this method to characterize the CP promoter collection that was BioBricked by the Uppsala 2013 team. |};
|
