Synthetic Gene DataBase
 

Synthetic Gene 134


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID124134
GenBank AccessionNC_001526
GenBank GI9627100
Gene NameE2E2 (16 E2-dm)
Gene Length (bp)10981098
Specieshuman papillomavirus (HPV-16)Homo sapiens
StrainsType 16human cervical cancer cell C33A
CDSatggagactctttgccaacgtttaaatgtgtgtcaggacaaaatactaacacattatgaa
aatgatagtacagacctacgtgaccatatagactattggaaacacatgcgcctagaatgt
gctatttattacaaggccagagaaatgggatttaaacatattaaccaccaagtggtgcca
acactggctgtatcaaagaataaagcattacaagcaattgaactgcaactaacgttagaa
acaatatataactcacaatatagtaatgaaaagtggacattacaagacgttagccttgaa
gtgtatttaactgcaccaacaggatgtataaaaaaacatggatatacagtggaagtgcag
tttgatggagacatatgcaatacaatgcattatacaaactggacacatatatatatttgt
gaagaagcatcagtaactgtggtagagggtcaagttgactattatggtttatattatgtt
catgaaggaatacgaacatattttgtgcagtttaaagatgatgcagaaaaatatagtaaa
aataaagtatgggaagttcatgcgggtggtcaggtaatattatgtcctacatctgtgttt
agcagcaacgaagtatcctctcctgaaattattaggcagcacttggccaaccaccccgcc
gcgacccataccaaagccgtcgccttgggcaccgaagaaacacagacgactatccagcga
ccaagatcagagccagacaccggaaacccctgccacaccactaagttgttgcacagagac
tcagtggacagtgctccaatcctcactgcatttaacagctcacacaaaggacggattaac
tgtaatagtaacactacacccatagtacatttaaaaggtgatgctaatactttaaaatgt
ttaagatatagatttaaaaagcattgtacattgtatactgcagtgtcgtctacatggcat
tggacaggacataatgtaaaacataaaagtgcaattgttacacttacatatgatagtgaa
tggcaacgtgaccaatttttgtctcaagttaaaataccaaaaactattacagtgtctact
ggatttatgtctatatga
atggagaccctgtgccagcgcctgaacgtgtgccaggacaagatcctgacccactacgag
aacgacagcaccgacctgcgcgaccacatcgactactggaagcacatgcgcctggcctgc
gccatctactacaaggcccgcgagatgggcttcaagcacatcaaccaccaggtggtgccc
accctggccgtgagcaagaacaaggccctgcaggccgccgagctgcagctgaccctggag
accatctacaacagccagtacagcaacgagaagtggaccctgcaggacgtgagcctggag
gtgtacctgaccgcccccaccggctgcatcaagaagcacggctacaccgtggaggtgcag
ttcgacggcgacatctgcaacaccatgcactacaccaactggacccacatctacatctgc
gaggaggccagcgtgaccgtggtggagggccaggtggactactacggcctgtactacgtg
cacgagggcatccgcacctacttcgtgcagttcaaggacgacgccgagaagtacagcaag
aacaaggtgtgggaggtgcacgccggcggccaggtgatcctgtgccccaccagcgtgttc
agcagcaacgaggtgagcagccccgagaccatccgccagcacctggccaaccacagcgcc
gccacccacaccaaggccgtggccctgggcaccgaggagacccagaccaccatccagcgc
ccccgcagcgagcccgacaccggcaacccctgccacaccaccaagctgctgcaccgcgac
agcgtggacagcgcccccatcctgaccgccttcaacagcagccacaagggccgcatcaac
tgcaacagcaacaccacccccatcgtgcacctgaagggcgacgccaacaccctgaagtgc
ctgcgctaccgcttcaagaagcactgcaagctgtacaccgccgtgagcagcacctggcac
tggaccggccacaacgtgaagcacaagagcgccatcgtgaccctgacctacgacagcgag
tggcagcgcgaccagttcctgagccaggtgaagatccccaagaccatcaccgtgagcacc
ggcttcatgagcatctaa
5' End
3' End
NotesSource for the natural gene was not provided in the paper. GenBank Accession# NC_001526 is used.Sequence from United States Patent & Trademark office, publication # 20050075303.
Expression VectorpV1J
Assay MethodsBeta-gal assay, Replication assay, Immunoblotting, Western Blot
ResultsLow.Retained substantial DNA replication function.
Protein Functionregulatory protein
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodRecoded by employing codons that are most commonly used by highly expressed mammalian genes.
Publication Author(s)Wang, X. M.; Jansen, K. U.; McClements, W. L.
Corresponding AuthorWilliam L. McClements
Corresponding AddressDepartment of Microbial Vaccine Research, P O Box 4, West Point, PA 19486, USA.
Publication Year2003
Publication TitleDNA replicative functions of highly-expressed, codon-optimized human papillomavirus proteins E1 and E2
AbstractHuman papillomavirus (HPV) DNA replication requires functional HPV early (E) proteins E1 and E2. To determine the biological activity of HPV 16 E1 and E2 mutant proteins under consideration as vaccine candidates, we developed a sensitive real time PCR assay that monitors HPV origin-of-replication-driven DNA synthesis. The assay was used to determine the DNA replicative functions of highly-expressed, codon-optimized HPV 16 E1 and E2 wild type and mutant proteins in transient transfections. Under the assay conditions, the HPV 16 E1 mutant (W439R, G482D) did not support HPV origin-driven DNA synthesis. In contrast, however, an HPV 16 E2 mutant bearing an E39A substitution, reported previously to be severely compromised for DNA replication, was found to be reduced only two-fold in activity and, therefore, considered not sufficiently inactivated for use in vaccines that depend on endogenous protein expression.
JournalJ Virol Methods. 108(1): 83-90.
SummaryTo determine the biological activity of HPV 16 E1 and E2 mutant proteins under consideration as vaccine candidates, codon-optimized HPV 16 E1 and E2 wild type and mutant genes were used. The codon optimized genes were constructed by employing codons that are most commonly used by highly expressed mammalian genes. The results showed that the HPV16 origin-driven DNA replication required both E1 and E2. The codon-optimized HPV16 E1 double mutant did not support any detectable DNA replication, but HPV 16 E2 single mutant as well as double mutant retained substantial DNA replication function. However, only codon-optimized wild type E1, together with codon-optimized wild type E2 support significant replication.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=626
PubMed ID12565157
Submitter NameZin, Htar
Submitter Address1000 Hilltop Circle, Baltimore, MD 21250
Entry ConfirmationNo
 
 

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