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Team:TU Munich/Project/Constitutive Promoter
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Here the sequences from YPA [[Media:Tum12Promotersequenzen.pdf]], [[Media:Tum12 Promoterliste.pdf]] | Here the sequences from YPA [[Media:Tum12Promotersequenzen.pdf]], [[Media:Tum12 Promoterliste.pdf]] | ||
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| - | == Vectorsystems for Yeast | + | == Vectorsystems for Yeast == |
A common commercial Vector system for Yeast transformation is the pESC lineage from Stratagene | A common commercial Vector system for Yeast transformation is the pESC lineage from Stratagene | ||
Revision as of 11:54, 15 August 2012
Contents |
BBa_K517001 GPD Constitutive Promoter
Background and principles
BBa_K517001 is a constitutive promoter. It was created by [British Columbia 2011]. They claim it is a commonly used promoter and they got it from [http://www.addgene.org/yeast-gateway/ Yeas Gateway].
Idea
This will be used for positive control for our constructs.
General remarks and issues
I could not find out what GPD stands for Functionality was shown for an induction time of 2 hours. The submitted DNA is faulty, so it probably needs fixing.
Biobricks and sequences
>BBa_K517001 Part-only sequence (112 bp)
cggtaggtattgattgtaattctgtaaatctatttcttaaacttcttaaattctacttttatagttagtcttttttttagttttaaaacaccagaactta gtttcgacggat
Assembly Compatibility:
10 12 21 23 25
| Library | Plate | Well | Plasmid Backbone | Resistance | |||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| 2011 Submission Plates | TP 4004 | 4G | pSB1C3 | ||||||||||||||||||
| |||||||||||||||||||||
References
http://partsregistry.org/wiki/index.php?title=Part:BBa_K517001 Partsregistry
Book reference: Christine Guthrie, Gerald R. Fink. 2004. Guide to Yeast Genetics and Molecular and Cell Biology, Volume 1. Gulf Professional Publishing. Chapter 26: Vectors for Constitutive and Inducible Gene Expression in Yeast. Page 391.
Non-Registry Promoters
Background and principles
Georg
Jie Sun et al. describes 14 constitutive promoters from Saccharomyces cerevisiae, and its reaction at different culture conditions, oxy(-)/glu(+),oxy(+)/glu(+). 2 Where described as higher strength promoters TEF1p, TPI1p, and 7 as medium strenth pr. GPM1p, GPDp (TDH3p), FBA1p, PDC1p, ENO2p, PYK1p, and TEF2p. With exception to the strongest TEF1p all exhibit nearly the same expression values with varying oxygen conditions. TEF1p was also tested for small glucose concentrations showed 100% expression levels with oxygen limitation and 70 % without oxygenlimitation after 80 hours of cell cultivation.
File:Geclonte promoter.jpg
A.Blount et al. states that repeated use of the same biological parts is not possible since the cells recombine long stretches of homologous DNA
Siavash Partow et al. compares several constitutive Promoters during varying Glucose concentrations Media:Tum12Promoterstärken.pdf, TEF1, HXT7,PGK1 are shown to be the most suitable ones. They also show the fusion of two promoters for bidirectional control of two genes namely TEF1 and PGK1 with comparable activity in both states Media:Tum12Promoterfusion.pdf, two constructs were successfully used pSP-G1-Vector and pSP-G2.
.
Media:Tum12Fusionspromoter_Vector.pdf
Elke Nevoigt et al. generated a library of TEF1 promotermutants for finetuning of Expression Media:Tum12 TEF-Promoter-mutants.pdf
Idea
General remarks and issues
TEF1p guarantues relative strong expression values under varying conditions as it is the case for brewing.
Biobricks and sequences
TEF1 sequence : from Promoter Database for Sacharomyces cerevisiae SCPD :(http://rulai.cshl.edu/cgi-bin/SCPD/getgenelist)
RAP1
GCCGTACCACTTCAAAACACCCAAGCACAGCATACTAAATTTCCCCTCTT -301
TCTTCCTCTAGGGTGTCGTTAATTACCCGTACTAAAGGTTTGGAAAAGAA -251
AAAAGAGACCGCCTCGTTTCTTTTTCTTCGTCGAAAAAGGCAATAAAAAT -201
TTTTATCACGTTTCTTTTTCTTGAAAATTTTTTTTTTTGATTTTTTTCTC -151
TTTCGATGACCTCCCATTGATATTTAAGTTAATAAACGGTCTTCAATTTC -101
TCAAGTTTCAGTTTCATTTTTCTTGTTCTATTACAACTTTTTTTACTTCT -51
TGCTCATTAGAAAGAAAGCATAGCAATCTAATCTAAGTTTTAATTACAAA
TPI1 Promotorsequenz
CTACTTATTCCCTTCGAGATTATATCTAGGAACCCATCAGGTTGGTGGAA -401
GCR1 RAP1
GATTACCCGTTCTAAGACTTTTCAGCTTCCTCTATTGATGTTACACCTGG -351
RAP1 GCR1
ACACCCCTTTTCTGGCATCCAGTTTTTAATCTTCAGTGGCATGTGAGATT -301
CTCCGAAATTAATTAAAGCAATCACACAATTCTCTCGGATACCACCTCGG -251
TTGAAACTGACAGGTGGTTTGTTACGCATGCTAATGCAAAGGAGCCTATA -201
TACCTTTGGCTCGGCTGCTGTAACAGGGAATATAAAGGGCAGCATAATTT -151
AGGAGTTTAGTGAACTTGCAACATTTACTATTTTCCCTTCTTACGTAAAT -101
ATTTTTCTTTTTAATTCTAAATCAATCTTTTTCAATTTTTTGTTTGTATT -51
CTTTTCTTGCTTAAATCTATAACTACAAAAAACACATACATAAACTAAAA -1
PGK1-Promoter
CGATTTGGGCGCGAATCCTTTATTTTGGCTTCACCCTCATACTATTATCA -601
REB1 ABF1
GGGCCAGAAAAAGGAAGTGTTTCCCTCCTTCTTGAATTGATGTTACCCTC -551
ABF1
ATAAAGCACGTGGCCTCTTATCGAGAAAGAAATTACCGTCGCTCGTGATT -501
RAP1 GCR1
TGTTTGCAAAAAGAACAAAACTGAAAAAACCCAGACACGCTCGACTTCCT -451
GTCTTCCTATTGATTGCAGCTTCCAATTTCGTCACACAACAAGGTCCTAG -401
CGACGGCTCACAGGTTTTGTAACAAGCAATCGAAGGTTCTGGAATGGCGG -351
GAAAGGGTTTAGTACCACATGCTATGATGCCCACTGTGATCTCCAGAGCA -301
AAGTTCGTTCGATCGTACTGTTACTCTCTCTCTTTCAAACAGAATTGTCC -251
GAATCGTGTGACAACAACAGCCTGTTCTCACACACTCTTTTCTTCTAACC -201
AAGGGGGTGGTTTAGTTTAGTAGAACCTCGTGAAACTTACATTTACATAT -151
ATATAAACTTGCATAAATTGGTCAATGCAAGAAATACATATTTGGTCTTT -101
TCTAATTCGTAGTTTTTCAAGTTCTTAGATGCTTTCTTTTTCTCTTTTTT -51
ACAGATCATCAAGGAAGTAATTATCTACTTTTTACAACAAATATAAAACA -1
For Yeast genes there is also another database calles yeast promoter atlas, the sequences however differ from the first one (http://ypa.ee.ncku.edu.tw/) Here the sequences from YPA Media:Tum12Promotersequenzen.pdf, Media:Tum12 Promoterliste.pdf
Vectorsystems for Yeast
A common commercial Vector system for Yeast transformation is the pESC lineage from Stratagene
a broad range of Protocols for yeast transformation and for the making of competent cells can be found on [http://www.protocol-online.org/prot/Model_Organisms/Yeast/Yeast_Transformation/index.html] from several laboratories that work with yeast.
Plasmids from Publications as well as from companies can be viewed at[http://www.addgene.org/] and can also be ordered there, as far as I could see they are far less expensive than those from companies.
