Difference between revisions of "Part:BBa K1741004"

Latest revision as of 07:31, 27 September 2015

sfGFP under melibiose promoter (improved 5'UTR)

Twice as high expression (comparing to BBa_K1741003) has been obtained by 5’UTR editing – removal of potential secondary structure which involved RBS and a better positioning of RBS – 7 nt upstream AUG start codon. The modified 5’UTR results in about twice higher expression upon melibiose addition to the M9 or 2xLB medium. The promoter is still weaker than comercial araBAD or rhaBAD promoters, so can be the promoter of choice if lower expression level is necessary for efficient folding or secretion of a recombinant protein.

Design

Legend:

MelS [BBa_K1741004]

MelWT [BBa_K1741003]

Results

Legend:

MelS [BBa_K1741004]
MelWT [BBa_K1741003]

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
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal SapI.rc site found at 207

@@ Line 1: / Line 1: @@
-Twice higher expression (comparing to BBa_K1741003) has been obtained by 5’UTR editing – removal of potential secondary structure which involved RBS and a better positioning of RBS – 7 nt upstream AUG start codon. The modified 5’UTR  results in about twice higher expression upon melibiose addition to the M9 or 2xLB medium. The promer is still weaker than araBAD or rhaBAD, so can be a promoter of choice if lower expression level is necessary for efficient folding or secretion of a recombinant protein.
+__NOTOC__
+<partinfo>BBa_K1741004 short</partinfo>
+Twice as high expression (comparing to BBa_K1741003) has been obtained by 5’UTR editing – removal of potential secondary structure which involved RBS and a better positioning of RBS – 7 nt upstream AUG start codon. The modified 5’UTR  results in about twice higher expression upon melibiose addition to the M9 or 2xLB medium. The promoter is still weaker than comercial araBAD or rhaBAD promoters, so can be the promoter of choice if lower expression level is necessary for efficient folding or secretion of a recombinant protein.
+===Design===
+Legend:
+MelS [[https://parts.igem.org/Part:BBa_K1741004 BBa_K1741004]]
+https://static.igem.org/mediawiki/parts/f/f1/UAMpoznanmelS.jpg
+MelWT [[https://parts.igem.org/Part:BBa_K1741003 BBa_K1741003]]
+https://static.igem.org/mediawiki/parts/0/02/UAMpoznanmelWT.jpg
+===Results===
+Legend:
+MelS [[https://parts.igem.org/Part:BBa_K1741004 BBa_K1741004]]
+<br>
+MelWT [[https://parts.igem.org/Part:BBa_K1741003 BBa_K1741003]]
+https://static.igem.org/mediawiki/parts/9/97/Teamuampoznanmelibiosem9graph.png
+https://static.igem.org/mediawiki/parts/2/2c/Teamuampoznanmelibioselbgraph.png
+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
+===Usage and Biology===
+<!-- -->
+<span class='h3bb'>Sequence and Features</span>
+<partinfo>BBa_K1741004 SequenceAndFeatures</partinfo>
+<!-- Uncomment this to enable Functional Parameter display
+===Functional Parameters===
+<partinfo>BBa_K1741004 parameters</partinfo>
+<!-- -->