Difference between revisions of "Part:BBa K1741004"
 
(5 intermediate revisions by 2 users not shown)
Line 1: Line 1:
 
__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K1741004 short</partinfo>
 
<partinfo>BBa_K1741004 short</partinfo>
−
 
−
==Design==
 
  
 
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.
 
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===
 +
  
  
Line 17: Line 18:
 
https://static.igem.org/mediawiki/parts/0/02/UAMpoznanmelWT.jpg
 
https://static.igem.org/mediawiki/parts/0/02/UAMpoznanmelWT.jpg
  
−
===Charecteristic and Results===
+
===Results===
  
 
Legend:
 
Legend:
Line 29: Line 30:
 
https://static.igem.org/mediawiki/parts/2/2c/Teamuampoznanmelibioselbgraph.png
 
https://static.igem.org/mediawiki/parts/2/2c/Teamuampoznanmelibioselbgraph.png
  
−
The  sfGFP fluorescence was measured using Tecan fluorometer.
+
The  sfGFP fluorescence [RFU] was measured using Tecan fluorometer.
  
 
We also checked the tightness of our promoters.
 
We also checked the tightness of our promoters.

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]

UAMpoznanmelS.jpg

MelWT [BBa_K1741003]

UAMpoznanmelWT.jpg

Results

Legend:

MelS [BBa_K1741004]
MelWT [BBa_K1741003]

Teamuampoznanmelibiosem9graph.png

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. Teamuampoznanpromoterscomparisongrpah.png


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