Generator

Part:BBa_K1741007

Designed by: Marta Żardecka   Group: iGEM15_UAM_Poznan   (2015-09-17)
Revision as of 06:07, 27 September 2015 by Daria (Talk | contribs)

sfGFP under xylose promoter xylA (xylWT)

SfGFP under xylose promoter xylA (wild type).

=Design

Legend:

XylWT (XylA) [BBa_K1741007]

XylA1 [BBa_K1741008]

XylS [BBa_K1741009]

XylF [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. 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.

Characteristics and Results

Legend:

XylWT (XylA) [BBa_K1741007]

XylA1 [BBa_K1741008]

XylS [BBa_K1741009]

XylF [BBa_K1741010]


Teamuampoznanxyloselbgraph.png

Teamuampoznanxylosem9graph.png

Teamuampoznanxyloseod600graph.png

The sfGFP fluorescence [RFU] was measured using Tecan fluoremeter.

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.

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 408


[edit]
Categories
Parameters
None