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Synthetic Gene 8


 
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Field NameNatural GeneSynthetic Gene
SGDB Gene ID88
GenBank Accession
GenBank GI0
Gene NameParathyroid hormone (PTH)PTH
Gene Length (bp)0
SpeciesHomo sapiensEscherichia coli
StrainsCAG629
CDS
5' End
3' End
Notes
Expression Vector
Assay Methods
Results20 fold increase
Protein FunctionEndosecretory hormone
Recoding PurposeTo improve expression
Synthesized ByAuthors
Recoding Method'(a) extending complementarity of the mRNA to the anticodon loop of tRNAfMet by use of a codon with
a purine nucleotide directly following the ATG, (b) avoidance of stable secondary structure in the
mRNA by use of synonymous A/U-rich codons, (c) elimination of unwanted restriction sites
Publication Author(s)Morelle, G.; Frank, R.; Meyerhans, A.
Corresponding AuthorAndreas Meyerhans
Corresponding AddressDivision of Enzymtechnology, GBF-Gesellschaft fur Biotechnologische Forschung, Braunschweig, F.R.G.
Publication Year1991
Publication TitleRestructuring the translation initiation region of the human parathyroid hormone gene for improved expression in Escherichia coli
AbstractOverexpression of native human parathyroid hormone in Escherichia coli was achieved by a modification of the 5' end of the genomic gene sequence, thereby adapting this part of the translation initiation region to the bacterial host. Some simple rules abstracted from optimization studies of translation initiation of a beta-interferon gene were applied. These included (a) extending complementarity of the mRNA to the anticodon loop of tRNAfMet by use of a codon with a purine nucleotide directly following the ATG, (b) avoidance of stable secondary structure in the mRNA by use of synonymous A/U-rich codons, (c) elimination of a potential second Shine-Dalgarno sequence. The appropriate silent changes led to a 20-fold increase in parathyroid hormone production resulting in 4.3% of total soluble protein. This result proves the validity of our simple approach for optimization of foreign gene expression in E. coli.
JournalBiochim Biophys Acta. 1089(3): 320-4.
SummaryThe goal of this experiment was to recode the translation initiation region on the hPTH gene to optimize expression in E. coli. The recoded gene was cloned into pJLA505 expression vector and then transformed into CAG629 strain E. coli. This resulted in a "remarkable increased expression" of PTH.
Comments
Discussion http://www.evolvingcode.net/forum/viewtopic.php?p=670#670
PubMed ID1859835
Submitter NameZheng, Yuanpu
Submitter AddressUMBC
Entry ConfirmationNo
 
 

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