Synthetic Gene DataBase

Synthetic Gene 110

  Welcome, Guest!

Field NameNatural GeneSynthetic Gene
SGDB Gene ID96110
GenBank AccessionAF287352
GenBank GI11066861
Gene Namegag-polgag-pol
Gene Length (bp)00
Specieshuman Immunodeficiency Virus (HIV-1)Homo sapiens
Strains293T, COS-1, NIH 3T3
5' End
3' End
NotesThe natural gene was not used in the experiments. Source is not provided.The authors provides this Accession number, which shows the synthetic constructs of gag and pol separately.
Expression VectorpcDNA3gagpolml
Assay MethodsWestern Blot, Statistical analysis
Resultsefficient expression
Protein Function
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding Methodused optimal codon usage for mammalian genes expression
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.
PubMed ID11747598
Submitter NameZin, Htar
Submitter Address1000 Hilltop Circle, Baltimore, MD 21250
Entry ConfirmationNo

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