Synthetic Gene DataBase
 

Synthetic Gene 113


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID102113
GenBank Accession
GenBank GI
Gene NamesbwAFPsynthetic sbwAFP
Gene Length (bp)0315
SpeciesChoristoneura fumiferanaNicotiana tabacum
StrainsLeaves
CDS
5' End
3' End
NotesNo accession number or CDS was provided for the natural gene or recoded gene. Corresponding author was contacted but unable to provide the information for the natural gene or the recoded gene.No accession number or CDS was provided for the natural gene or recoded gene. Corresponding author was contacted but unable to provide the information for the natural gene or the recoded gene.
Expression VectorpBI121 with CaMV 35s promoter
Assay MethodsSDS-PAGE
ResultsSignificant increase (amount comparable to naturally occurring AFP type III)
Protein Function
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodConstruction of recoded gene was based on the codon usage of highly expressed genes in tobacco
plants. Codons with C and G in the second and third positions were removed and as well as ones with
T and A in the second and third positions. PR terminal leader sequence was fused to the frame of the
gene as well.
Publication Author(s)Holmberg, N.; Farres, J.; Bailey, J. E.; Kallio, P. T.
Corresponding AuthorPauli T. Kallio
Corresponding AddressUnilever Research, Colworth House, Sharnbrook, Bedford MK44 1LQ, UK.
Publication Year2001
Publication TitleTargeted expression of a synthetic codon optimized gene, encoding the spruce budworm antifreeze protein, leads to accumulation of antifreeze activity in the apoplasts of transgenic tobacco
AbstractA synthetic gene based on the primary sequence of the mature spruce budworm antifreeze protein (sbwAFP) was constructed by primer overlap extension. The amino acid codons were chosen to mimic those of a highly expressed tobacco nuclear gene. A DNA sequence encoding the amino-terminal leader sequence from the tobacco pathogen related protein 1b (PR), which targets the protein to the apoplastic space, was fused in frame to the synthetic sbwAFP gene. This fusion was placed downstream of the cauliflower mosaic virus 35S promoter and upstream of the nopaline synthase terminator in a T-DNA binary vector. Transgenic tobacco lines transcribing PR-sbwAFP were selected by RT-PCR. The apoplastic protein fractions of sbwAFP expressing tobacco lines exhibited enhanced antifreeze activity as demonstrated by the ability to inhibit ice re-crystallization and increased thermal hysteresis.
JournalGene. 275(1): 115-24.
SummaryTobacco plants (Nicotiana tabacum) were transformed with synthetic sbwAFP, a recoded version of the sbwAFP gene, in order to increase the expression of the antifreeze protein. sbwAFP had its codons modified based on the codon usage of highly expressed tobacco plant genes in order to get the recoded gene synthetic sbwAFP. Synthetic sbwAFP was able to produce a significant amount of protein capable of being used for its intended purpose. The recoded gene was successful overall in its goal to provide an increase in protein expression.
CommentsNo accession number or CDS was provided for the natural gene or recoded gene. Corresponding author was contacted but unable to provide the information for the natural gene or the recoded gene.
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=564
PubMed ID11574159
Submitter NameQureshi, Imran
Submitter Address1000 Hilltop Circle Baltimore, MD 21250 USA
Entry ConfirmationNo
 
 

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