Difference between revisions of "Part:BBa K1170000:Experience"
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<I>BABS UNSW 2015</I> | <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_K1033225">BBa_K1033225</a>) and ASR (<a href="https://parts.igem.org/Part:BBa_K1170000">BBa_K1170000) 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> | + | <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_K1033225">BBa_K1033225</a>) and ASR (<a>href="https://parts.igem.org/Part:BBa_K1170000">BBa_K1170000</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> | <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> | ||
Latest revision as of 18:42, 18 September 2015
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Applications of BBa_K1170000
BABS UNSW 2015
A LacZ assay was carried out with p170 (BBa_K1677300), CP44 (BBa_K1033225) and ASR (href="https://parts.igem.org/Part:BBa_K1170000">BBa_K1170000) 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.
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UNIQ197527b84d9465cc-partinfo-00000001-QINU UNIQ197527b84d9465cc-partinfo-00000002-QINU