Difference between revisions of "Part:BBa K1741008"

Revision as of 06:13, 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 our XylA1 promoter we have modified the 5'UTR of the XylWT promoter so it was similar to that of proD [BBa_K1741014], a strong constitutive promoter. In some experiments it seems to be slightly stronger than original XylA promoter from part BBa_K1741007. The next step was to shorten the promoter by removing its XylF part while keeping the modified UTR. Finally we tested the XylF promoter upstream of sfGFP. XylS is the strongest xylose-induced promoter. Xylose induced promoter XylA1 is weaker than arabinose and rhamnose promoters.

Characteristics and 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 fluoremeter.

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==
+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 our XylA1 promoter we have modified the 5'UTR of the XylWT promoter so it was similar to that of proD [BBa_K1741014], a strong constitutive promoter. In some experiments it seems to be slightly stronger than original XylA promoter from part BBa_K1741007.
+The next step was to shorten the promoter by removing its XylF part while keeping the modified UTR. Finally we tested the XylF promoter upstream of sfGFP. XylS is the strongest xylose-induced promoter.
+Xylose induced promoter XylA1 is weaker than arabinose and rhamnose promoters.
 ===Characteristics and Results===
-https://static.igem.org/mediawiki/parts/0/0a/Teamuampoznanxyloseod600graph.png
+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/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 fluoremeter.
 <!-- Add more about the biology of this part here