Difference between revisions of "Part:BBa K1677300:Experience"

Latest revision as of 18:16, 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_K1677300

A LacZ assay was carried out with p170, CP44 and ASR 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.
Overall this suggests that our biobricked p170 works as predicted by Madsen et al. (1999) and that it will up-regulate transcription at lower pH values.
Notably this is a significant result as the assay was carried out in E. Coli, not L. Lactis, where the promoter was initially characterized.

User Reviews

UNIQ4f19c7290eb8f207-partinfo-00000001-QINU UNIQ4f19c7290eb8f207-partinfo-00000002-QINU

@@ Line 5: / Line 5: @@
 ===Applications of BBa_K1677300===
 <html>
-<br>A LacZ assay was carried out with p170, CP44 and ASR fused to a promoter activity reporter (BBa_I732095) 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, CP44 and ASR 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> 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> Overall this suggests that our biobricked p170 works as predicted by Madsen et al. (1999) and that it will up-regulate transcription at lower pH values. <br> Notably this is a significant result as the assay was carried out in E. Coli, not L. Lactis, where the promoter was initially 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>