Difference between revisions of "Part:BBa K801074:Design"

Latest revision as of 10:30, 24 October 2012

CaMXMT1 expression cassette for yeast

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1433
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 205

Composite part of BBa_K319003, BBa_K801071 and BBa_K801012

Keywords: caffeine, TEF1, expression cassette

Abbreviations:

CaMXMT1 = coffea arabica 7-methylxanthine N-methyltransferase 1

Design Notes

Related BioBrick: BBa_K801071

Quality control measures:

Test digest has been performed with use of Xba1 and Pst1 (see mainpage)

Backbone:

Backbone name: pSB1C3'
Resistance: Cp

Protein coding:

Protein: CaMXMT1-Strep-tag

Enzymatic activity:

EC-number [http://enzyme.expasy.org/EC/2.1.1.160 2.1.1.160]

Cytotoxicity:
This Biobrick is one part of the pathway, which leads to caffeine (the BioBricks BBa_801073 and BBa_801075 are further needed). 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_K801071

Organism:

Genesequence derived from Coffea arabica
codonoptimized for Saccharomyces cerevisiae
designed for Saccharomyces cerevisiae

References

Literature references:

[http://www.ncbi.nlm.nih.gov/pubmed/12746542 Pubmed: 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.]
[http://www.ncbi.nlm.nih.gov/pubmed/16247553 Pubmed: Uefuji H., Ogita S., Yamaguchi Y., Koizumi N. and Sano H., 2005: Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant.]
[http://www.ncbi.nlm.nih.gov/pubmed/16925551 Pubmed: Kuranda K, Leberre V, Sokol S, Palamarczyk G, François J., 2006: Investigating the caffeine effects in the yeast Saccharomyces cerevisiae brings new insights into the connection between TOR, PKC and Ras/cAMP signalling pathways.]

Database references:

NCBI

@@ Line 14: / Line 14: @@
 <!--*used_abbreviation_1 = full_name_of_used_abbreviations_1-->
 <!--*used_abbreviation_2 = full_name_of_used_abbreviations_2-->
+* CaMXMT1 = coffea arabica 7-methylxanthine N-methyltransferase 1
 ===Design Notes===
@@ Line 91: / Line 92: @@
 '''Literature references:'''<br>
+*[http://www.ncbi.nlm.nih.gov/pubmed/12746542 '''Pubmed:''' 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.]
+*[http://www.ncbi.nlm.nih.gov/pubmed/16247553 '''Pubmed:''' Uefuji H., Ogita S., Yamaguchi Y., Koizumi N. and Sano H., 2005: Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant.]
+*[http://www.ncbi.nlm.nih.gov/pubmed/16925551 '''Pubmed:'''  Kuranda K, Leberre V, Sokol S, Palamarczyk G, François J., 2006: Investigating the caffeine effects in the yeast Saccharomyces cerevisiae brings new insights into the connection between TOR, PKC and Ras/cAMP signalling pathways.]
 <!--*[http://www.ncbi.nlm.nih.gov/pubmed/?PMID? '''Pubmed:''' ?Author(s)?, ?year?: ?title?]-->
 '''Database references:'''<br>
+* NCBI
 <!--*[http://www.ncbi.nlm.nih.gov/nuccore/?accessNr? '''GenBank''': ?title?]-->
 <!--*[http://www.ebi.ac.uk/interpro/IEntry?ac=?accessNr? '''Interpro''': ?title?]-->