Synthetic Gene DataBase
 

Synthetic Gene 147


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID134147
GenBank AccessionAY177679AJ313179
GenBank GI2775286015212100
Gene NameL1oriL1h
Gene Length (bp)15181518
Specieshuman papillomavirus (HPV-16)Homo sapiens, Mus musculus
StrainsType 16293T, 911 cells
CDSatgtctctttggctgcctagtgaggccactgtctacttgcctcctgtcccagtatctaag
gttgtaagcacggatgaatatgttgcacgcacaaacatatattatcatgcagggacatcc
agactacttgcagttggacatccctattttcctattaaaaaacctaacaataacaaaata
ttagttcctaaagtatcaggattacaatacagggtatttagaatacatttacctgacccc
aataagtttggttttcctgacacctcattttataatccagatacacagcggctggtttgg
gcctgtgtaggtgttgaggtaggccgtggtcagccattaggtgtgggcattagtggccat
cctttattaaataaattggatgacacagaaaatgctagtgcttatgcagcaaatgcaggt
gtggataatagagaatgtatatctatggattacaaacaaacacaattgtgtttaattggt
tgcaaaccacctataggggaacactggggcaaaggatccccatgtaccaatgttgcagta
aatccaggtgattgtccaccattagagttaataaacacagttattcaggatggtgatatg
gttgatactggctttggtgctatggactttactacattacaggctaacaaaagtgaagtt
ccactggatatttgtacatctatttgcaaatatccagattatattaaaatggtgtcagaa
ccatatggcgacagcttatttttttatttacggagggaacaaatgtttgttagacattta
tttaatagggctggtgctgttggtgaaaatgtaccagacgatttatacattaaaggctct
gggtctactgcaaatttagccagttcaaattattttcctacacctagtggttctatggtt
acctctgatgcccaaatattcaataaaccttattggttacaacgagcacagggccacaat
aatggcatttgttggggtaaccaactatttgttactgttgttgatactacacgcagtaca
aatatgtcattatgtgctgccatatctacttcagaaactacatataaaaatactaacttt
aaggagtacctacgacatggggaggaatatgatttacagtttatttttcaactgtgcaaa
ataaccttaactgcagacgttatgacatacatacattctatgaattccactattttggag
gactggaattttggtctacaacctcccccaggaggcacactagaagatacttataggttt
gtaacatcccaggcaattgcttgtcaaaaacatacacctccagcacctaaagaagatccc
cttaaaaaatacactttttgggaagtaaatttaaaggaaaagttttctgcagacctagat
cagtttcctttaggacgcaaatttttactacaagcaggattgaaggccaaaccaaaattt
acattaggaaaacgaaaagctacacccaccacctcatctacctctacaactgctaaacgc
aaaaaacgtaagctgtaa
atgagcctgtggctgcccagcgaggccaccgtgtacctgccccccgtgcccgtgagcaag
gtggtgagcaccgacgagtacgtggccaggaccaacatctactaccacgccggcaccagc
aggctgctggccgtgggccacccctacttccccatcaagaagcccaacaacaacaagatc
ctggtgcccaaggtgagcggcctgcagtacagggtgttcaggatccacctgcccgacccc
aacaagttcggcttccccgacaccagcttctacaaccccgacacccagaggctggtgtgg
gcctgcgtgggcgtggaggtgggcaggggccagcccctgggcgtgggcatcagcggccac
cccctgctgaacaagctggacgacaccgagaacgccagcgcctacgccgccaacgccggc
gtggacaacagggagtgcatcagcatggactacaagcagacccagctgtgcctgatcggc
tgcaagccccccatcggcgagcactggggcaagggcagcccctgcaccaacgtggccgtg
aaccccggcgactgcccccccctggagctgatcaacaccgtgatccaggacggcgacatg
gtggacaccggcttcggcgccatggacttcaccaccctgcaggccaacaagagcgaggtg
cccctggacatctgcaccagcatctgcaagtaccccgactacatcaagatggtgagcgag
ccctacggcgacagcctgttcttctacctgaggagggagcagatgttcgtgaggcacctg
ttcaacagggccggcgccgtgggcgagaacgtgcccgacgacctgtacatcaagggcagc
ggcagcaccgccaacctggccagcagcaactacttccccacccccagcggcagcatggtg
accagcgacgcccagatcttcaacaagccctactggctgcagagggcccagggccacaac
aacggcatctgctggggcaaccagctgttcgtgaccgtggtggacaccaccaggagcacc
aacatgagcctgtgcgccgccatcagcaccagcgagaccacctacaagaacaccaacttc
aaggagtacctgaggcacggcgaggagtacgacctgcagttcatcttccagctgtgcaag
atcaccctgaccgccgacgtgatgacctacatccacagcatgaacagcaccatcctggag
gactggaacttcggcctgcagcccccccccggcggcaccctggaggacacctacaggttc
gtgaccagccaggccatcgcctgccagaagcacaccccccccgcccccaaggaggacccc
ctgaagaagtacaccttctgggaggtgaacctgaaggagaagttcagcgccgacctggac
cagttccccctgggcaggaagttcctgctgcaggccggcctgaaggccaagcccaagttc
accctgggcaagaggaaggccacccccaccaccagcagcaccagcaccaccgccaagagg
aagaagaggaagctgtga
5' End
3' End
NotesGenbank accession # for native gene is not provided in the paper. AY177679 is used for the record.GenBank Accession No. is provided by the authors.
Expression VectorpUF3pUF3
Assay MethodsELISA, Western Blot, Electron microscopy, Nuclear staining, Flow cytometry.ELISA, Western Blot, Electron microscopy, Nuclear staining, Flow cytometry.
ResultsUndetectable.Significant increased.
Protein Functionmajor capsid protein
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding Methodcodon usage adopted for expression in human cells according to Kazuka codon
usage database
Publication Author(s)Leder, C.; Kleinschmidt, J. A.; Wiethe, C.; Muller, M.
Corresponding AuthorMartin Muller
Corresponding AddressForschungsschwerpunkt fur Angewandte Tumorvirologie, Deutsches Krebsforschungszentrum Heidelberg, 69120 Heidelberg, Germany.
Publication Year2001
Publication TitleEnhancement of capsid gene expression: preparing the human papillomavirus type 16 major structural gene L1 for DNA vaccination purposes
AbstractExpression of the structural proteins L1 and L2 of the human papillomaviruses (HPV) is tightly regulated. As a consequence, attempts to express these prime-candidate genes for prophylactic vaccination against papillomavirus-associated diseases in mammalian cells by means of simple DNA transfections result in insufficient production of the viral antigens. Similarly, in vivo DNA vaccination using HPV L1 or L2 expression constructs produces only weak immune responses. In this study we demonstrate that transient expression of the HPV type 16 L1 and L2 proteins can be highly improved by changing the RNA coding sequence, resulting in the accumulation of significant amounts of virus-like particles in the nuclei of transfected cells. Data presented indicate that, in the case of L1, adaptation for codon usage accounts for the vast majority of the improvement in protein expression, whereas translation-independent posttranscriptional events contribute only to a minor degree. Finally, the adapted L1 genes demonstrate strongly increased immunogenicity in vivo compared to that of unmodified L1 genes.
JournalJ Virol. 75(19): 9201-9.
SummaryThe goal of this study is to overcome the inefficient expression of HPV-16 L1 for vaccination purposes in cells which do not resemble differentiating keratinocytes. For this purpose, the synthetic HPV-16 L1 genes in which the codon usage was optimized for plant cells, Solanum tuberosum L1p, or mammalian cells, H. sapiens L1h, were constructed. In both constructs, the majority of the codons were modified while the encoded protein sequence remained unchanged. The experimental results showed that the codon optimized L1 genes increased the expression level and immunogenecity significantly compared to that of noncodon-optimized L1 genes.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=655
PubMed ID11533183
Submitter NameZin, Htar
Submitter Address1000 Hilltop Circle, Baltimore, MD 21250
Entry ConfirmationNo
 
 

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