Synthetic Gene DataBase
 

Synthetic Gene 21


 
  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID1921
GenBank AccessionAF123610
GenBank GI10442764
Gene NameTAR1TAR1 R
Gene Length (bp)20222022
SpeciesTriticum aestivumSaccharomyces cerevisiae
StrainsClone RED1-TAWAT11
CDSatgacgtcgcatgcacgcgtacgtaagcttggatcctctagagcggccgcgctcctggcc
gggctgctcgtgctcgccttctggctccgatccggcgggagcgcgccgcccaagccgccc
cccgcgtccttccgcccgccgcccgtcaaggtcgacgccgacgacgccgacgacggccgc
aagcgcgtcaccatcttcttcggcacccagaccggcaccgccgagggcttcgccaaggct
atggcggaggaggccaaggcgcggtacgagaaggtcctcttcaaagtggtggatctggat
gactatgctgccgaggacgatgagtacgaggagaagctcaagaaggaaacttttgcgctc
ttcttcttagcaacatatggcgacggggaacccactgataatgcagcaagattctacaaa
tggttcactgagggtaaggagaaggaggtttggcttaaggatttcaattatgctgtattt
ggccttggcaacaggcaatacgagcattttaataaggttgcaaaggaggtggatgagtta
ctgctggagcagggtggcaatcgtcttgttccatgtggccttggtgatgatgatcagtgc
attgaggatgacttcaccgcatggaaagaactagtatggccagaattggaccaactgctc
cgtgatgaagatgatactactggtgcaaccactccttacactgctgcaatacctgaatat
agagttgtatttattgataaatcagatttagagttcgaagataaatcctggacacttgcc
aacggcaacggtgttattgatgcccaacacccttgcagggccaatgttgctgttcggaag
gaactccataaaccagcctcggatcgctcttgcattcatctggaatttgatatttcaggc
actggtcttgtgtatgaaactggggatcatgttggggtttattcagaaaattctatcgag
acagtggagcaggctgaaacgctacttgatctttccccagatacagttttctccatacat
gcagacgccgaagatggatctcctcgtaaaggcgggggttccttggctccaccattccca
tctccttgtaccttacggactgcacttctacgatatgctgatctactcaatgctccaaag
aaggctgctttgactgctttagctgctcatgcttctgacccgactgaagctgagcgactg
aggttcttggcttctcctgctggaaaggatgagtatgctcaatggatagtagctagccag
aggagccttcttgaagttatggctgcattcccttcagctaagccccccttgggggtattc
tttgcggcgatagctcctcgtctacagccacgattctactccatatcatcatctccaaag
ttggctccctccagaattcatgtgacatgtgcattagtttatggtcctacaccaacagaa
aggatccaccaaggagtttgttcgacatggatgaagaacacacttccattggaatatagc
gaagagtgcagttgggcaccaatttttgtgaggcagtcaaattttaggttgcctgcagat
ccatccactccaatcatcatgattggtcctgggacagggctagcaccttttaggggcttc
ttgcaggaaagattagctctaaaagaatctggagctgaattgggctcgtcagtcctcttc
tttggatgcaggaaccgtgaaatggactacatatatgaggaagagttgcaaaatttcctt
caggagggggcactctctgagctaattgtcgcctactctcgtgaaggaccaaccaaagaa
tatgtgcagcataagatggtggacaaggccaccgagatttggaacgtcatttctcaaggt
ggttacatatatgtttgtggtgatgccaagggcatggcgaaagacgtacacagggttctc
cacacaatagttcaaaaacagggatctttggataactccaaaactgagctgtatgtaaag
aacctacagatggaaggcaggtacctccgtgatgtgtggtaa
atgacttcacatgctagagttagaaaattaggttcttctagagctgcagctttattggct
ggtttattggttttagctttttggttaagatctggtggtagtgctcctcctaaaccaccc
cccgcgtccttccgcccgccgcccgtcaaggtcgacgccgacgacgccgacgacggccgc
aagcgcgtcaccatcttcttcggcacccagaccggcaccgccgagggcttcgccaaggct
atggcggaggaggccaaggcgcggtacgagaaggtcctcttcaaagtggtggatctggat
gactatgctgccgaggacgatgagtacgaggagaagctcaagaaggaaacttttgcgctc
ttcttcttagcaacatatggcgacggggaacccactgataatgcagcaagattctacaaa
tggttcactgagggtaaggagaaggaggtttggcttaaggatttcaattatgctgtattt
ggccttggcaacaggcaatacgagcattttaataaggttgcaaaggaggtggatgagtta
ctgctggagcagggtggcaatcgtcttgttccatgtggccttggtgatgatgatcagtgc
attgaggatgacttcaccgcatggaaagaactagtatggccagaattggaccaactgctc
cgtgatgaagatgatactactggtgcaaccactccttacactgctgcaatacctgaatat
agagttgtatttattgataaatcagatttagagttcgaagataaatcctggacacttgcc
aacggcaacggtgttattgatgcccaacacccttgcagggccaatgttgctgttcggaag
gaactccataaaccagcctcggatcgctcttgcattcatctggaatttgatatttcaggc
actggtcttgtgtatgaaactggggatcatgttggggtttattcagaaaattctatcgag
acagtggagcaggctgaaacgctacttgatctttccccagatacagttttctccatacat
gcagacgccgaagatggatctcctcgtaaaggcgggggttccttggctccaccattccca
tctccttgtaccttacggactgcacttctacgatatgctgatctactcaatgctccaaag
aaggctgctttgactgctttagctgctcatgcttctgacccgactgaagctgagcgactg
aggttcttggcttctcctgctggaaaggatgagtatgctcaatggatagtagctagccag
aggagccttcttgaagttatggctgcattcccttcagctaagccccccttgggggtattc
tttgcggcgatagctcctcgtctacagccacgattctactccatatcatcatctccaaag
ttggctccctccagaattcatgtgacatgtgcattagtttatggtcctacaccaacagaa
aggatccaccaaggagtttgttcgacatggatgaagaacacacttccattggaatatagc
gaagagtgcagttgggcaccaatttttgtgaggcagtcaaattttaggttgcctgcagat
ccatccactccaatcatcatgattggtcctgggacagggctagcaccttttaggggcttc
ttgcaggaaagattagctctaaaagaatctggagctgaattgggctcgtcagtcctcttc
tttggatgcaggaaccgtgaaatggactacatatatgaggaagagttgcaaaatttcctt
caggagggggcactctctgagctaattgtcgcctactctcgtgaaggaccaaccaaagaa
tatgtgcagcataagatggtggacaaggccaccgagatttggaacgtcatttctcaaggt
ggttacatatatgtttgtggtgatgccaagggcatggcgaaagacgtacacagggttctc
cacacaatagttcaaaaacagggatctttggataactccaaaactgagctgtatgtaaag
aacctacagatggaaggcaggtacctccgtgatgtgtggtaa
5' Endcggcccgggccatcgat
3' Endgagctcgggagctcgg
Notes
Expression VectorpYeDP110pYeDP110
Assay MethodsP450 CO-binding spectraEnzymatic activity measurements
ResultsUndetectableDramatic increase in the enzymaic activity
Protein FunctionEnzyme: NADPH-cytochrome P450 reductase
Recoding PurposeTo improve expression
Synthesized ByAuthor
Recoding MethodFirst 39 codons were optimized with reference to yeast highly expressed genes (Sharp and Cowe,
1991).
Publication Author(s)Batard, Y.; Hehn, A.; Nedelkina, S.; Schalk, M.; Pallett, K.; Schaller, H.; Werck-Reichhart, D.
Corresponding AuthorDaniele Werck-Reichhart
Corresponding AddressDepartement d'Enzymologie Cellulaire et Moleculaire, Institut de Biologie Moleculaire des Plantes, Centre National de la Recherche Scientifique FRE 2161, 28 rue Goethe, Strasbourg Cedex, 67083, France.
Publication Year2000
Publication TitleIncreasing expression of P450 and P450-reductase proteins from monocots in heterologous systems
AbstractMonocotyledonous crop plants are usually more resistant to herbicides than grass weeds and most dicots. Their resistance to herbicides is mediated in many cases by P450 oxygenases. Monocots thus constitute an appealing source of P450 enzymes for manipulating herbicide resistance and recombinant forms of the major xenobiotic metabolizing mooxygenases are potential tools for the optimization of new active molecules. We report here the isolation and functional characterization of the first P450 and P450 reductase coding sequences from wheat. The first attempts at expressing these cDNAs in yeast and tobacco led to levels of protein, which were extremely low, often not even detectable. The wheat P450 cDNAs were efficiently transcribed, but no protein or activity was found. Wheat coding sequences, like those of other monocots, are characterized by a high GC content and by a related strong bias of codon usage, different from that observed in yeast or dicots. Complete recoding of genes being costly, the reengineering their 5'-end using a single PCR megaprimer designed to comply with codon usage of the host was attempted. It was sufficient to relieve translation inhibition and to obtain good levels of protein expression. The same strategy also resulted in a dramatic increase in protein expression in tobacco. A basis for the success of such a partial recoding strategy, much easier and cheaper than complete recoding of the cDNA, is proposed.
JournalArch Biochem Biophys. 379(1): 161-9.
SummaryThe N-terminal part of three wheat genes were optimized with reference to the the codon usage in highly expressed genes. As measured by the P450 CO-binding essay, the codon optimization increased the protein expression. However, the expression seems to be only slightly improved when looking at the western blot results.
CommentsThe first 17 codons of TAR1 gene in the paper were not showed in GenBank but have been manually added
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=518
PubMed ID10864454
Submitter NameWu, Gang
Submitter AddressDepartment of Biological Sciences, University of Maryland Baltimore County, 1000 Hilltop Circle, Baltimore, MD 21250 USA
Entry ConfirmationNo
 
 

Copyright 2004 the Freeland Bioinformatics Lab, All Rights Reserved. | Contact Us | About this site