Team:Hong Kong HKUST/pneumosensor/module two
From 2014.igem.org
Line 120: | Line 120: | ||
<div class= "content_area_one_row"> | <div class= "content_area_one_row"> | ||
<p> | <p> | ||
- | + | ||
Combox promoters can be found on many different regions within the genomic DNA of Streptococcus Pneumoniae strains. The promoters were reported to have variants with different lengths and consensus sequences, but generally the range of variety has been kept small. Though much information about the combox promoter is readily available nowadays, its characterization of promoter activity, specificity, sequence, as well as the biomolecular mechanism can be greatly enhanced with further investigations and experiments.<br><br> | Combox promoters can be found on many different regions within the genomic DNA of Streptococcus Pneumoniae strains. The promoters were reported to have variants with different lengths and consensus sequences, but generally the range of variety has been kept small. Though much information about the combox promoter is readily available nowadays, its characterization of promoter activity, specificity, sequence, as well as the biomolecular mechanism can be greatly enhanced with further investigations and experiments.<br><br> | ||
Line 135: | Line 135: | ||
</table> | </table> | ||
+ | <!--two row of content, image first row, text second row --> | ||
+ | <div class='content_1'><h3> Reporting Module Description </h3> | ||
<table class="content_table" align= "center" > | <table class="content_table" align= "center" > | ||
- | + | ||
- | <td class= "content_cell"> | + | <tr class= "content_row" > |
+ | <td class= "content_cell" colspan= "2"> | ||
<div class= "content_area_one_row"> | <div class= "content_area_one_row"> | ||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
- | |||
<div class="content_image"> | <div class="content_image"> | ||
- | <img src= "https://static.igem.org/mediawiki/2014/ | + | <img src= "https://static.igem.org/mediawiki/2014/f/fd/Diagram_text_2_edited_copy.png"/> |
</div> | </div> | ||
- | + | <p>Besides comX, another positive factor involved in competence regulation was later found out to be ComW. The gene comW (SP0018) is regulated by the quorum-sensing system and is required for a high-level of competence. Coexpression of comW with comX restores the accumulation of σx and the expression of late genes as ComW contributes to the stabilization of the alternative sigma factor σx against proteolysis by ClpXP and is required for full activity of σx in directing transcription of late competence genes. | |
- | <p> Besides | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
</p> | </p> | ||
- | + | </div> | |
- | <p>Based on these findings, we | + | </td> |
- | + | <td class= "content_cell" colspan= "2"> | |
- | + | <div class= "content_area_one_row"> | |
- | + | <p> | |
- | + | ||
+ | |||
+ | Based on these findings, we integrated this alternative sigma factor system from Gram-positive Streptococcus pneumoniae into Gram-negative Escherichia coli. We firstly cloned out the comX and comW genes from the genomic DNA of S. pneumoniae NCTC 7465 strain. We then used BBa_K880005 (consisting of constitutive promoter J23100 and strong RBS B0034) from the BioBricks to express those genes.<br><br> | ||
+ | |||
+ | Lastly, we combined these constructs with combox promoter and GFP generator to check the functionality of the system, and calculate the relative promoter unit of combox promoter. | ||
+ | |||
+ | |||
+ | |||
</p> | </p> | ||
+ | |||
</div> | </div> | ||
</td> | </td> | ||
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
<div class='content_1'> | <div class='content_1'> |
Revision as of 14:47, 3 October 2014
Reporting Module Description
Prof. Donald A. Morrison’s research lab in University of Illinois at Chicago published several papers on the competence for genetic transformation in Streptococcus pneumoniae which depends on quorum-sensing system to control many competence-specific genes acting in DNA uptake, processing, and integration. There is a link between this quorum-sensing system and the competence-specific genes, which is an alternative sigma factor ComX, σx. Expression of ComX allows transcription of many genes that are involved in transformation and specifically expressed during competence. These late genes share a conserved 8-bp sequence in their promoter regions, TACGAATA (combox) which is specifically induced by σx-containing RNA polymerase. |
Combox promoters can be found on many different regions within the genomic DNA of Streptococcus Pneumoniae strains. The promoters were reported to have variants with different lengths and consensus sequences, but generally the range of variety has been kept small. Though much information about the combox promoter is readily available nowadays, its characterization of promoter activity, specificity, sequence, as well as the biomolecular mechanism can be greatly enhanced with further investigations and experiments. |
Reporting Module Description
Besides comX, another positive factor involved in competence regulation was later found out to be ComW. The gene comW (SP0018) is regulated by the quorum-sensing system and is required for a high-level of competence. Coexpression of comW with comX restores the accumulation of σx and the expression of late genes as ComW contributes to the stabilization of the alternative sigma factor σx against proteolysis by ClpXP and is required for full activity of σx in directing transcription of late competence genes. |
Based on these findings, we integrated this alternative sigma factor system from Gram-positive Streptococcus pneumoniae into Gram-negative Escherichia coli. We firstly cloned out the comX and comW genes from the genomic DNA of S. pneumoniae NCTC 7465 strain. We then used BBa_K880005 (consisting of constitutive promoter J23100 and strong RBS B0034) from the BioBricks to express those genes. |
References
|
Home |
Pneumosensor |
Riboregulator |
Human Practice |
Team |
WetLab |
Achievement |