Synthetic Gene DataBase
 

Synthetic Gene 109


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID95109
GenBank AccessionAF287355
GenBank GI11066868
Gene Namepolpol
Gene Length (bp)03009
Specieshuman Immunodeficiency Virus (HIV-1)Homo sapiens
Strains293T, COS-1, NIH 3T3
CDSatgagggaagatctggccttcccacaagggaaggccaggaaattttcttcagagcagacc
agagccaacagccccatcagaagagagcgtcaggtttggagaagagacaacaactccctc
tcagaagcaggagccgatagacaaggaactgtatcctttagcttccctcagatcactctt
tggcagcgacccctcgtcacaataaagatcggcggccagctgaaggaggccctgctggac
accggcgccgacgacaccgtgctggaggagatgaacctgcccggcaggtggaagcccaag
atgatcggcggcatcggcggcttcatcaaggtgaggcagtacgaccagatccccatcgag
atctgcggccacaaggccatcggcaccgtgctggtgggccccacccccgtgaacatcatc
ggcaggaacctgctgacccagatcggctgcaccctgaacttccccatcagccccatcgag
accgtgcccgtgaagctgaagcccggcatggacggccccaaggtgaagcagtggcccctg
accgaggagaagatcaaggccctggtggagatctgcaccgagatggagaaggagggcaag
atcagcaagatcggccccgagaacccctacaacacccccgtgttcgccatcaagaagaag
gacagcaccaagtggaggaagctggtggacttcagggagctgaacaagaggacccaggac
ttctgggaggtgcagctgggcatcccccaccccgccggcctgaagaagaagaagagcgtg
accgtgctggacgtgggcgacgcctacttcagcgtgcccctgcacgaggacttcaggaag
tacaccgccttcaccatccccagcatcaacaacgagacccccggcaccaggtaccagtac
aacgtgctgccccagggctggaagggcagccccgccatcttccagagcagcatgaccacc
atcctggagcccttcaggaagcagaaccccgacctggtgatctaccagtacatggacgac
ctgtacgtgggcagcgacctggagatcggccagcacaggaccaagatcgaggagctgagg
cagcacctgctgaggtggggcttcaccacccccgacaagaagcaccagaaggagcccccc
ttcctgtggatgggctacgagctgcaccccgacaagtggaccgtgcagcccatcgtgctg
cccgagaaggacagctggaccgtgaacgacatccagaagctggtgggcaagctgaactgg
gccagccagatctacgccggcatcaaggtgaggcagctgtgcaagctgctgaggggcacc
aaggccctgaccgaggtgatccccctgaccgaggaggccgagctggagctggccgagaac
agggagatcctgaaggagcccgtgcacggcgtgtactacgaccccagcaaggacctgatc
gccgagatccagaagcagggccagggccagtggacctaccagatctaccaggagcccttc
aagaacctgaagaccggcaagtacgccaggaccaggggcgcccacaccaacgacgtgaag
cagctgaccgaggccgtgcagaagatcgccaccgagagcatcgtgatctggggcaagacc
cccaagttcaagctgcccatccagaaggagacctgggagacctggtggaccgagtactgg
caggccacctggatccccgagtgggagttcgtgaacaccccccccctggtgaagctgtgg
taccagctggagaaggagcccatcatcggcgccgagaccttctacgtggacggcgccgcc
aacagggagaccaagctgggcaaggccggctacgtgaccaacaagggcaggcagaaggtg
gtgagcctgaccgacaccaccaaccagaagaccgagctgcaggccatctacctggccctg
caggacagcggcctggaggtgaacatcgtgaccgacagccagtacgccctgggcatcatc
caggcccagcccgacaggagcgagagcgagctggtgagccagatcatcgagcagctgatc
aagaaggagaaggtgtacctggcctgggtgcccgcccacaagggcatcggcggcaacgag
caggtggacaagctggtgagcgccggcatcaggaaggtgctgttcctggacggcatcgac
aaggcccaggaggagcacgagaagtaccacagcaactggagggccatggccagcgacttc
aacctgccccccgtggtggccaaggagatcgtggccagctgcgacaagtgccagctgaag
ggcgaggccatgcacggccaggtggactgcagccccggcatctggcagctggactgcacc
cacctggagggcaaggtgatcctggtggccgtgcacgtggccagcggctacatcgaggcc
gaggtgatccccgccgagaccggccaggagaccgcctacttcctgctgaagctggccggc
aggtggcccgtgaccaccatccacaccgacaacggcagcaacttcaccagcgccaccgtg
aaggccgcctgctggtgggccggcatcaagcaggagttcggcatcccctacaacccccag
agccagggcgtggtggagagcatgaacaaggagctgaagaagatcatcggccaggtgagg
gaccaggccgagcacctgaagaccgccgtgcagatggccgtgttcatccacaacttcaag
aggaagggcggcatcggcggctacagcgccggcgagaggatcgtggacatcatcgccacc
gacatccagaccaaggagctgcagaagcagatcaccaagatccagaacttcagggtgtac
tacagggacagcagggaccccctgtggaagggccccgccaagctgctgtggaagggcgag
ggcgccgtggtgatccaggacaacagcgacatcaaggtggtgcccaggaggaaggccaag
atcatcagggactacggcaagcagatggccggcgacgactgcgtggccggcaggcaggac
gaggactag
5' End
3' End
NotesThe natural gene was not used in the experiments. Source is not provided
Expression VectorpcDNApolml
Assay MethodsWestern Blot, Statistical analysis
Resultsnot efficient expression.
Protein Function
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding Method
Publication Author(s)Fuller, M.; Anson, D. S.
Corresponding AuthorD.S. Anson
Corresponding AddressDepartment of Chemical Pathology, Women's and Children's Hospital, North Adelaide South Australia, 5006.
Publication Year2001
Publication TitleHelper plasmids for production of HIV-1-derived vectors
AbstractVectors derived from human immunodeficiency virus type 1 (HIV-1) appear an attractive option for many gene therapy applications. This is due to their ability to transduce noncycling cell populations and to integrate their genome into the host cell chromosome, resulting in the stable genetic modification of the transduced cell. These properties have permitted the direct in vivo transduction of several tissues, including the central nervous system, retina, and liver. However, the pathogenic nature of HIV-1 has raised considerable concerns about the safety of such vector systems. To help address these concerns, we have expressed each of the primary transcriptional units encoding trans functions relevant for vector production in individual plasmid constructs. The gag-pol gene sequence was codon-optimized for expression in mammalian cells resulting in high level Rev/Rev-response element (RRE)-independent expression. Codon optimization of gag-pol also reduces sequence homology with vectors containing gag gene sequences, which results in reduced transfer of biologically active gag-pol sequences to transduced cells. Furthermore, the vif reading frame overlapping the 3' end of the pol coding sequence is destroyed by codon optimization. We have also shown that the Gag and Gag-Pol polyproteins can be efficiently expressed from separate transcriptional units. This has enabled the removal of a cis-acting viral element, the gag-pol translational frameshift sequence, from the vector/packaging system and prevents detectable transfer of biologically active sequences equivalent to the gag-pol gene to transduced cells.
JournalHum Gene Ther. 12(17): 2081-93.
SummaryTo facilitate the production of recombinant virus for gene therapy studies with increased safety, a series of constructs were made for the expression of HIV1 proteins and combined with a suitable vectors. The synthetic constructs for gag-pol(gagpolml), gag (gagml), and pol (polml) genes were made based on the optimal codon usage for mammalian gene expression. The variant of the synthetic gag-pol sequence, gagpolfusionml , in which the frameshift of the region of overlap between the reading frames encoding gag pol were replaced by codon- optimized sequence encoding gag-pol polyprotein. According to the experimental results, codon-optimized gag-pol gene sequence (pcDNA3gagpolml) showed expression and virus production efficiently, the expressions of codon optimized gag (pcDN3gagml), pol (pcDNA3polm), and pcDNA3gagpolfusionml on their own can not support the production of virus however. In contrast, the combination of the two plasmids pcDN3gagm and pcDNA3gagpolfusionml can replace the pcDNA3gagpolml codon-optimized construct.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=661
PubMed ID11747598
Submitter NameZin, Htar
Submitter Address1000 Hilltop Circle, Baltimore, MD 21250
Entry ConfirmationNo
 
 

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