Difference between revisions of "Part:BBa K1741008"

Latest revision as of 08:52, 27 September 2015

sfGFP under promoter xylA1 with improved 5'UTR

Design

Legend:

Genes involved in catabolism of xylose in E.coli are organized into two transcription units which are driven by two promoters - XylA and XylF. In order to increase the expression of a protein under control of our XylA1 promoter we added the 5'UTR of the proD promoter[BBa_K1741014] to the XylA of XylWT. In some experiments it seems to be slightly stronger than the original XylA promoter from part BBa_K1741007. The next step was to shorten the promoter by removing its XylF part while keeping the modified 5'UTR (from proD) behind XylA. Finally we tested the reversed XylF part of the XylWT promoter with sfGFP as the reporter gene. XylS is the strongest xylose-induced promoter. Xylose induced promoter XylA1 is weaker than arabinose and rhamnose promoters.

Results

Legend:

XylWT (XylA) [BBa_K1741007]

XylA1 [BBa_K1741008]

XylS [BBa_K1741009]

XylF [BBa_K1741010]

All our xylose promoters are induced by xylose and repressed by glucose. We also observed slight induction by arabinose.

We observed that XylA1 and XylF had a lower expression than XylWT. XylS is the strongest xylose-induced promoter. The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.

We also checked the tightness of our promoters.

All of our promoters are induced only by their respective sugars with a small exception of xylose promoters being slightly induced by arabinose and arabinose promoters being slightly induced by xylose. All of our promoters are repressed by glucose.

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 153
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 419

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1741008 short</partinfo>
-Xylose induced promoter xylA is weaker than arabinose and  rhamnose promoters. To enhance the efficiency of xylA driven transcription/translation we have transplanted 5’UTR from the strong constitutive promoter proD. In some experiments it seems to be slightly stronger than original xylA promoter from part BBa_K1741007.
+===Design===
-===Characteristics and Results===
+Legend:
+XylWT (XylA) [[https://parts.igem.org/Part:BBa_K1741007 BBa_K1741007]]
+XylA1 [[https://parts.igem.org/Part:BBa_K1741008 BBa_K1741008]]
+XylS [[https://parts.igem.org/Part:BBa_K1741009 BBa_K1741009]]
+XylF [[https://parts.igem.org/Part:BBa_K1741010 BBa_K1741010]]
+Genes involved in catabolism of xylose in ''E.coli'' are organized into two transcription units which are driven by two promoters - XylA and XylF. In order to increase the expression of a protein under control of our XylA1 promoter we added the 5'UTR of the proD promoter[BBa_K1741014] to the XylA of XylWT. In some experiments it seems to be slightly stronger than the original XylA promoter from part BBa_K1741007. The next step was to shorten the promoter by removing its XylF part while keeping the modified 5'UTR (from proD) behind XylA. Finally we tested the reversed XylF part of the XylWT promoter with sfGFP as the reporter gene.
+XylS is the strongest xylose-induced promoter. Xylose induced promoter XylA1 is weaker than arabinose and rhamnose promoters.
+===Results===
+Legend:
+XylWT (XylA) [[https://parts.igem.org/Part:BBa_K1741007 BBa_K1741007]]
+XylA1 [[https://parts.igem.org/Part:BBa_K1741008 BBa_K1741008]]
+XylS [[https://parts.igem.org/Part:BBa_K1741009 BBa_K1741009]]
+XylF [[https://parts.igem.org/Part:BBa_K1741010 BBa_K1741010]]
-https://static.igem.org/mediawiki/parts/0/0a/Teamuampoznanxyloseod600graph.png
 https://static.igem.org/mediawiki/parts/f/f8/Teamuampoznanxyloselbgraph.png
 https://static.igem.org/mediawiki/parts/5/5d/Teamuampoznanxylosem9graph.png
+https://static.igem.org/mediawiki/parts/0/0a/Teamuampoznanxyloseod600graph.png
+All our xylose promoters are induced by xylose and repressed by glucose. We also observed slight induction by arabinose.
+We observed that XylA1 and XylF had a lower expression than XylWT. XylS is the strongest xylose-induced promoter.
+The sfGFP fluorescence [RFU] was measured using Tecan fluorometer.
+We also checked the tightness of our promoters.
+All of our promoters are induced only by their respective sugars with a small exception of xylose promoters being slightly induced by arabinose and arabinose promoters being slightly induced by xylose. All of our promoters are repressed by glucose.
+https://static.igem.org/mediawiki/parts/4/43/Teamuampoznanpromoterscomparisongrpah.png
 <!-- Add more about the biology of this part here