Synthetic Gene DataBase

Synthetic Gene 246

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Field NameNatural GeneSynthetic Gene
SGDB Gene ID218246
GenBank AccessionAY750900
GenBank GI53987910
Gene NameLLO91A (listeriolysin O aa 91-99)LLO91E
Gene Length (bp)3030
SpeciesListeria monocytogenesMus musculus; African Green Monkey
StrainsBALB/3T3; COS-7
5' End
3' End
Notessequence taken from Uchijima 1998 (same gene construct)
Expression VectorpLLO91A-LucpLLO91E-Luc
Assay Methodsluciferase activityluciferase activity
ResultsThe relative luciferase activity was 10^2.The relative luciferase activity was 10^4.
Protein FunctionH-2Kd- restricted CTL epitope
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodRecoding progressively increase the CAI value Mus musculus and decreased the CAI values for the
source species.
Publication Author(s)Nagata, T.; Uchijima, M.; Yoshida, A.; Kawashima, M.; Koide, Y.
Corresponding AuthorToshi Nagata
Corresponding AddressDepartment of Microbiology and Immunology, Hamamatsu University School of Medicine, 3600 Handa-cho, Hamamatsu, 431-3192, Japan.
Publication Year1999
Publication TitleCodon optimization effect on translational efficiency of DNA vaccine in mammalian cells: analysis of plasmid DNA encoding a CTL epitope derived from microorganisms
AbstractInterspecific difference of codon usage is one of the major obstacles for effective induction of specific immune responses against bacteria and protozoa by DNA immunization. Using genes encoding major histocompatibility complex class I-restricted cytotoxic T-lymphocyte (CTL) epitopes, derived from an intracellular bacterium, Listeria monocytogenes and a mouse malaria parasite, Plasmodium yoelii, we report here that the codon optimization level of the genes is not precisely proportional to, but does correlate well with the translational efficiency in mammalian cells, which is concomitantly associated with the induction level of specific CTL response in the mouse. These results suggest that DNA immunization using the gene codon-optimized to mammals through the entire region is very effective.
JournalBiochem Biophys Res Commun. 261(2): 445-51.
SummaryThe authors wished to test the correlation between the gene sequences coding for certain MHC class I restricted CTL epitopes and their relative expression in mammalian cells, and ultimately the level of immune response in mice. The genes for listeriolysin O (amino acids 91-99) and circumsporozoite protein (aa 281-290) were recoded so that codon usage progressive resembled a typical mammalianized gene. RSCU and CAI values were shown to track the progression. A total of 5 genes were synthesized for LLO and 4 genes for CSP. The epitopes were tagged with luciferase and relative levels increased with increasingly aggressive recoding. LLO’s relative luciferase activity increased about 100-fold from LLO91-A to LLO91E. CSP’s expression did not improve so drastically, increasing less than 10-fold. Both LLO’s and CSP’s correlation between recoding and expression levels were positively correlated. In addition, CSP’s CAI value and luciferase activity were almost directly proportional.
PubMed ID10425204
Submitter NameZheng, Yuanpu
Submitter AddressUMBC
Entry ConfirmationNo

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