Difference between revisions of "Part:BBa K801077:Design"

(→‎References)
 
(4 intermediate revisions by the same user not shown)
Line 9: Line 9:
 
<br>'''Keywords:'''
 
<br>'''Keywords:'''
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
 
<!--These keywords are necessary to find your part using a fulltext sarch.-->
−
caffeine, TEF1,  
+
caffeine, TEF1, TEF2, ADH1, expression cassette
  
 
<br>'''Abbreviations:'''
 
<br>'''Abbreviations:'''
 
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
 
<!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
 
<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
 
<!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
 +
* CaXMT1 = coffea arabica xanthosine N-methyltransferase 1
 +
 +
* CaMXMT1 = coffea arabica 7-methylxanthine N-methyltransferase 1
 +
 +
* CaDXMT1 = coffea arabica 3,7-dimethylxanthine N-methyltransferase 1
  
 
===Design Notes===
 
===Design Notes===
Line 26: Line 31:
  
 
'''Cloning details:'''<br>
 
'''Cloning details:'''<br>
 +
* cloning in RFC10
 
<!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part-->
 
<!--*Designed in RFC10/RFC23/RFC25/RFC25_N-part-->
 
<!--*Mutation C889G to delete XbaI restriction site-->
 
<!--*Mutation C889G to delete XbaI restriction site-->

Latest revision as of 10:26, 24 October 2012

Caffeine Synthesis Pathway pTEF2-XMT1-tADH1-pTEF1-MXMT1-tADH1-pTEF2-DXMT1-tADH1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3613
    Illegal BglII site found at 5829
    Illegal BglII site found at 5925
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 138
    Illegal NgoMIV site found at 4317
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 2385


Composite part of BBa_K801010 BBa_K801070 BBa_K801012 BBa_K319003 BBa_K801071 BBa_K801012 BBa_K801010 BBa_K801072 BBa_K801012


Keywords: caffeine, TEF1, TEF2, ADH1, expression cassette


Abbreviations:

  • CaXMT1 = coffea arabica xanthosine N-methyltransferase 1
  • CaMXMT1 = coffea arabica 7-methylxanthine N-methyltransferase 1
  • CaDXMT1 = coffea arabica 3,7-dimethylxanthine N-methyltransferase 1

Design Notes

Related BioBrick: BBa_K801073 BBa_K801074 BBa_K801075 BBa_K801076

Quality control measures:
Test digest has been performed with use of Xba1 and Pst1 (see mainpage) Part hasn´t been sequenced

Backbone:

  • Backbone name: pSB1C3'
  • Resistance: Cp

Protein coding:

  • Protein: CaXMT1-Strep-tag, CaMXTM1-Strep-tag, CaDXMT1-Strep-tag

Enzymatic activity:

  • see above related biobricks

Cytotoxicity:
This Biobrick includes all enzymes of the pathway which leads to caffeine. We investigated the toxicity of caffeine and found out, that the growth of yeast strains was inhibited at concentrations of 10 mM. Caffeine with a concentration of 100 mM even showed a lethal effect on yeast cells. Lower concentrations of caffeine (range of micro molar) did not show any growth inhibition or effect.

Safety notes:

Intellectual property:

Corresponding part author/authors:

Source

Source:

  • Preexisting BioBrick BBa_K801073,BBa_K801074, BBa_K801075


Organism:

  • Genesequence derived from coffea arabica
  • codonoptimized for saccharomyces cerevisiae
  • Designed for the following Chassis: saccharomyces cerevisiae

References

Literature references:
Uefuji, H., Ogita, S., Yamaguchi, Y., Koizumi, N., and Sano, H. (2003). Molecular cloning and functional characterization of three distinct n-methyltransferases involved in the caffeine biosynthetic pathway in coffee plants. Plant Physiol, 132(1):372–80.

Database references:

  • NCBI