Synthetic Gene DataBase
 

Synthetic Gene 254


 
  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID224254
GenBank AccessionAF367411
GenBank GI26557006
Gene NameSgp (SIV gag-pol)Sgpsyn
Gene Length (bp)43461
SpeciesSIVmnd-2Homo sapiens
StrainsT-REx 293, HeLa
CDSatgggcgcgagcgcgtcaggtcttaggggagaaaaattggacgagctggaaaagattagg
ttacggccctccggaaaaaagaaataccagctgaaacacataatatgggtaagcaaggaa
ctagacagatttggccttcatgaaaagctgttagaaagtaaggaaggatgcgagaaaatt
cttagcgtactctttccactagttcctacagggtcagaaaatttaatctcgctgtacaac
acctgctgctgcgtatggtgcgtacatgcgaaagagaaagtaacagatacagaggaagcg
aaagagaaagtaaaacaaaagctccatctagtggccgaaaaggaaaatgcagcatcagaa
aaagaacaaagagcaatagtgacacctagtggccgctcaaaaaattacccaatacagata
ataaatcagaccccagtccatcagggaatttcaccgcgcacgctgaatgcgtgggtaaaa
tgcatagaagagaagaagtttagcccagaaatagtgcccatgttcatagccttgtctgaa
gggtgcctcccctatgatctcaatggcatgctcaatgctattggagaccatcagggagcg
cttcaaatagtgaaggatgtcatcaatgaggaagctgcagactggaacttgagacaccca
caagtgggtcctatgccccaaggtgtcttgagaaacccaacgggcagtgatattgctgga
acaacaagctccatagaagagcagatagaatggactacaagggagcaagatgcagtaaat
gtgggaggaatatataaacaatggatagttttagggcttcagaagtgtgtaagcatgtat
aatccagtaaacatcctggacataaagcaaggaccaaaggagccgttcaaggactatgtg
gacaggttttacaaagctctgcgggcggagcggactgatccacaagtcaaaacctggatg
acacaaactttgctcatccagaatgctaacccagattgtaaatccatccttaaagggcta
ggcatgaatccctctttagaagagatgccgctagcctgccaaggggtaggaggccctaaa
tataaagcacaaatgatggcagaggctatgaaggaagcccagtcagctgtaatgatgcag
aattcgggagggccaccgcggggccccccgagacaacctcctagaaacatcagatgccct
aactgtggaaaatttggtcatgggctgagggactgtataagccctcgaaaaaagggatgc
ttcaagtgtggagatctaggacacataatgagaaattgcccaaagatggtgaatttttta
gggaataccccctggggcagtggcaaacccaggaacttcccggcgatgccattgacccca
acggcccctccaatgccagggatggaagacccagcagagagaatgctgttagattacatg
aagaaagggcaacagcagaaagcggaaagcaaacaggaaaagaaagagaggggtccatac
gaggcagcttacaactccctcagttctctctttggaacagaccaactacagtagtagaga
tagaagggcaaaaagtggaggccttactggacacaggagctgatgatacagtaatcaaag
atctagatttaaaaggtaattggaaaccacagattattggaggaattggaggatcaatca
atgtaaaacagtttttcaattgtaaagtaacaatagcaggcaaaactacacacgcttcag
tcctagtgggccccacacctgtaaatattgtaggtagaaatgtgctgaagaaattaggat
gtacactaaattttccagttagtaaagtagaaacagtaaaggtaacactaaaaccaggaa
ctgacggacctaaaataaaacaatggccattgtctaaggaaaagattttagccttacaag
aaatatgtagtcaaatggagaaggaaggccagatctctaagataggtccagaaaatcctt
acaacacaccggtgttttgcatcaaaaagaaagatggaaccagctggagaaaattggtag
attttagacagttaaacaaagtaactcaggacttctttgaggtgcagttgggaatcccac
accctggaggtctcaaacaatgtgagcaaattacagtactggacataggggatgcctatt
tctcatgtcctttggatgaggactttagaaagtacactgcattcaccattccatcggtga
ataatcagggcccaggaatcaggtaccagtataatgtcctaccacagggatggaagggct
ccccagccattttccaggcaacagcagataaaatcttacagcccttcagagagagacatc
cagatgtagtgatctatcaatatatggatgatctctttgttgggagtgatagagttgccc
cagaacacagcagaatgattcaagagttaagagaccacctcttgttttgggggctcgaga
ccccagacaagaagtttcaaaaggaaccaccctttgagtggatgggatacatactgcacc
ctaagaaatggacagtgcagaaagtacagcttccagagaaagaagaatggacagtaaatg
acatccaaaagttggtagggaaacttaattgggcaagtcagatttattccggaattaaaa
caaaagagctctgtagactaattagaggggcaaaacccctagatgaaaaagtagaatgga
ctagagaagcagaattagaatatgaagaaaacaaactcatagtgcaggaggaagtgcatg
gagtatactatcaaccagagaagcctttaatggcaaaggttcaaaaattgacacaagggc
aatggagttaccaaatagagcaggaggataataagccgctgaaggtagggaaatatgcca
gaacaaagaatgctcatacaaatgagcttagagtgcttgcaggactagtacagaagattg
caaaagaagctttagtgatctggggaaaactccctaagttttatctgccaatagaaagag
aagtctgggaccagtggtggccagaatattggcaagccacatggattcccgaatgggaat
ttgtatccactccacaccttatcgggttgtggtataacctgttgagagaacctgtgccag
gagaagatgtgtattatgtggatggagcagctaacagaaactccaaagaagggaaagcag
gatatgtaacagctagaaataaatccagagtaatagccctagagaataccacaaaccaaa
aagcagaattggaggcaataaaaatggcattacaagactcaggaccaaaagtaaatatag
tgacagattcacagtatgctatgggtatcctatcggcagcacccgaccaatcagacaacc
ccatagtaagagaaattatagaactcatgatccacaaggaagcagtatacctagcttggg
taccagcacacaaaggcataggagggaatgagcaagtagataaactagtgagcagaggag
taaggcaagtactgtttttagaaggcatagataaagcacaagaagagcatgataaatatc
acaataattggagggcattagcacaagacttctgtataccaaacatagtggcaaaagaaa
tagtagcccagtgtccaaagtgtcaaacaaaaggagagccaatacatggccaggtagata
catccccaggaacctggcaaatggactgcacacatatggaaggaaaagtaatcatagcag
cagtccatgtagcaagtaggtatctagaagcagaagtaatacccacagaaacaggaaaag
aaacagcacacttcctgttaaaattagcaggtagatggccagtaaaacatctacacacag
ataatggccctaatttcaccagtgaaaaagtagccacagtctgttggtgggctcaaatag
agcacactacaggaattccctacaatcctcagagtcaaggagtaatagaggcaaaaaacc
atcatctcaaacaaatcatagggcaagttagagaccaagcagagaaactagaaacagcag
ttcaaatggcagtattaattcacaattttaaaagaaaaggggggataggggaatacagtc
caggagaaagaatagtagacattatagcaacagacctcctaacaactaaattacaacaca
atattcaaaaaattcaaaattttcgggtttattacagagaaggaagggaccaacagtgga
aaggaccagcagaactcatttggaaaggagaaggagcggtggtaatcaaagaggggacag
acttgaaggtagtaccaaggagaaaagcaaaaatcatcagagattatggaaaaacagtgg
atagtgatcccaacgtggaagcttga
5' Endggccgatcgatgttgacattgattattgac
3' Endatgcggtgggctctatgggctagcagct
NotesWild-type genes were not tested in this paper.Emailed corresponding author for sequence 2/14/2006.
Expression VectorpShuttle-Sgpsyn
Assay MethodsSouthern Blot, Western Blot, Lentiviral titer
ResultsIn the presence of tetracycline, vector titers were about 100x higher than (-)Tet. Titers were on the order of 10^5 GFU/ml. SIV gag-pol expression was slightly lower than HIV gag-pol.
Protein Functionstructural genes
Recoding PurposeTo improve expression
Synthesized ByGeneart
Recoding Method“Codon-optimized versions of gag-pol were used, which reduces the risk of homologous recombination
and transfer of packing genes to target cells.”
Publication Author(s)Kuate, S.; Stefanou, D.; Hoffmann, D.; Wildner, O.; Uberla, K.
Corresponding AuthorKlaus Uberla
Corresponding AddressDepartment of Molecular and Medical Virology, Ruhr-University Bochum, D-44780 Bochum, Germany.
Publication Year2004
Publication TitleProduction of lentiviral vectors by transient expression of minimal packaging genes from recombinant adenoviruses
AbstractBACKGROUND: The potential of lentiviral vectors for clinical gene therapy has not yet been evaluated. One of the reasons is the cytotoxicity of lentiviral packaging genes which makes the generation of stable producer cell lines difficult. Therefore, a novel packaging system for lentiviral vectors based on transient expression of packaging genes by recombinant adenoviruses was developed. METHODS: Adenoviral vectors expressing VSV-G, codon-optimized HIV-1 gag-pol, and codon-optimized SIV gag-pol under the control of a tetracycline-regulatable promoter (adenoviral lenti-pack vectors) were constructed and the production levels of this vector system were evaluated. RESULTS: The generated adenoviral lenti-pack vectors could be grown to high titers when transgene expression was suppressed and no evidence for instabilities was obtained. Cells stably transfected with a SIV-based vector construct were converted into lentiviral vector producer cells by infection with the adenoviral lenti-pack vectors. Lentiviral vector titers obtained were as high as vector titers obtained by transient cotransfection experiments. A protocol was developed that allowed preparation of lentiviral vector stocks with undetectable levels of contaminating adenoviral lenti-pack vectors. CONCLUSIONS: The adenoviral lenti-pack vectors described should provide a convenient alternative approach to inducible packaging cell lines for large-scale lentiviral vector production. Transient expression of cytotoxic lentiviral packaging genes by the adenoviral lenti-pack vectors circumvents loss of titers during prolonged culture of packaging cell lines. The design of the adenoviral lenti-pack vectors should reduce the risk of transfer of packaging genes to target cells and at the same time provide flexibility with respect to the lentiviral vector constructs that can be packaged.
JournalJ Gene Med. 6(11): 1197-205.
SummaryThe authors wish to recode SIV and HIV genes in order to reduce the chance of packaging genes being transferred with genes of interest, the risk being a recombinant replication competent lentivirus. VSV-G, HIV-1 gag-pol, and SIV gag-pol were placed under the control of tetracycline-regulatable promoters and transfected into producer cells (T-REx 293), which would generate lentiviral vector particles that could enter target cells (HeLa). The authors did not make any mention of how the gene were recoded, simply that it was done by Geneart. Assays showed that vector production was strictly under the control of the tetracycline promoter.
CommentsThis paper is primarily about a gene transfer system and not codon optimization.
Discussion
PubMed ID15459964
Submitter NameZheng, Yuanpu
Submitter AddressUMBC
Entry ConfirmationNo
 
 

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