Synthetic Gene DataBase
 

Synthetic Gene 94


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID8194
GenBank AccessionM62653
GenBank GI155660
Gene NameGFPGFP
Gene Length (bp)7170
SpeciesAequorea victoriaHomo sapiens
StrainsCOS cells
CDSatgagtaaaggagaagaacttttcactggagttgtcccaattcttgttgaattagatggt
gatgttaatgggcacaaattttctgtcagtggagagggtgaaggtgatgcaacatacgga
aaacttacccttaaatttatttgcactactggaaaactacctgttccatggccaacactt
gtcactactttctcttatggtgttcaatgcttttcaagatacccagatcatatgaaacag
catgactttttcaagagtgccatgcccgaaggttatgtacaggaaagaactatatttttc
aaagatgacgggaactacaagacacgtgctgaagtcaagtttgaaggtgatacccttgtt
aatagaatcgagttaaaaggtattgattttaaagaagatggaaacattcttggacacaaa
ttggaatacaactataactcacacaatgtatacatcatggcagacaaacaaaagaatgga
atcaaagttaacttcaaaattagacacaacattgaagatggaagcgttcaactagcagac
cattatcaacaaaatactccaattggcgatggccctgtccttttaccagacaaccattac
ctgtccacacaatctgccctttcgaaagatcccaacgaaaagagagaccacatggtcctt
cttgagtttgtaacagctgctgggattacaca
5' End
3' End
NotesSource of the wld type GFP gene was not specified in the paper. Therefore, the M62653 was collected.
Expression Vector
Assay Methods
ResultsUndetectable.There was a net improvement in fluorescence per cell of between 40-120 fold, depending on detection condition.
Protein FunctionReporter gene
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding MethodThe systematic replacement of the native codons of GFP with codons chosen to reflect more closely
the codon preference of highly expressed human genes.The initiation consensus was replaced with
sequences corresponding to the translational initiation consensus.
Publication Author(s)Haas, J.; Park, E. C.; Seed, B.
Corresponding AuthorBrian Seed
Corresponding AddressDepartment of Molecular Biology, Massachusetts General Hospital, Boston, Massachusetts 02114, USA.
Publication Year1996
Publication TitleCodon usage limitation in the expression of HIV-1 envelope glycoprotein
AbstractBACKGROUND. The expression of both the env and gag gene products of human immunodeficiency virus type 1 (HIV-1) is known to be limited by cis elements in the viral RNA that impede egress from the nucleus and reduce the efficiency of translation. Identifying these elements has proven difficult, as they appear to be disseminated throughout the viral genome. RESULTS. Here, we report that selective codon usage appears to account for a substantial fraction of the inefficiency of viral protein synthesis, independent of any effect on improved nuclear export. The codon usage effect is not specific to transcripts of HIV-1 origin. Re-engineering the coding sequence of a model protein (Thy-1) with the most prevalent HIV-1 codons significantly impairs Thy-1 expression, whereas altering the coding sequence of the jellyfish green fluorescent protein gene to conform to the favored codons of highly expressed human proteins results in a substantial increase in expression efficiency. CONCLUSIONS. Codon-usage effects are a major impediment to the efficient expression of HIV-1 genes. Although mammalian genes do not show as profound a bias as do Escherichia coli genes, other proteins that are poorly expressed in mammalian cells can benefit from codon re-engineering.
JournalCurr Biol. 6(3): 315-24.
SummaryTo explore whether codon bias accounted for the poor expression of envelope glycoproteins, a synthetic gene encoding the gp120 segment of HIV1 based on the sequence of the prototype virus of the most common North American subtype, HIV-1MN. The native codons of gp120 were replaced with codon chosen to reflect more closely the codon preference of highly expressed human genes. Comparisons were made on the expression of wild-type and synthetic gp120. The synthetic gene product was expressed at a very high level compared to that of the wild-type gp120. To explore futher the importance of codon usage in envelope protein expression, synthetic rat Thy-1 gene and synthetic GFP gene of the jellyfish were constructed. The codons of wild-type Thy-1 were replaced with the codons most frequrntly used by the HIV-1 envelope protein. The synthetic GFP gene was created as the same manner as synthetic gp120. Codon replacement enhanced green fluorescent protein expression. However, HIV1 codon pattern undermine the efficiency of Thy-1 expression.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?t=671
PubMed ID8805248
Submitter NameZin, Htar
Submitter Address1000 Hilltop Circle, Baltimore, MD 21250
Entry ConfirmationNo
 
 

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