Chemical synthesis and expression of a gene coding for human muscle acylphosphatase

Alessandra Modesti, Giovanni Raugei, Niccolo' Taddei, Riccardo Marzocchini, Manuela Vecchi, Guido Camici, Giampaolo Manao, Giampietro Ramponi

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

A DNA sequence coding for human muscle acylphosphatase has been constructed using 16 chemically synthesized oligonucleotides. The 300-bases long DNA sequence has been cloned in the pT7.7 Escherichia coli expression vector and in the pYEpsec1 Saccharomyces cerevisiae expression vector. In both cases a high level of expression of acylphosphatase has been observed. The recombinant proteins have been purified to homogeneity and assayed in comparison with the natural protein, using benzoylphosphate as a substrate and phosphate as a competitive inhibitor. The recombinant enzymes expressed in the two microorganisms maintain the kinetic properties of the natural protein. In addition, NMR analysis shows that the gross fold of the two recombinant enzymes is correct.

Original languageEnglish
Pages (from-to)369-374
Number of pages6
JournalBBA - Gene Structure and Expression
Volume1216
Issue number3
DOIs
Publication statusPublished - Dec 14 1993

Fingerprint

DNA sequences
Muscle
Genes
Gene Expression
Muscles
Enzymes
Recombinant Proteins
Oligonucleotides
Microorganisms
Yeast
Escherichia coli
Saccharomyces cerevisiae
Proteins
Phosphates
Nuclear magnetic resonance
Kinetics
Substrates
acylphosphatase

Keywords

  • (Human)
  • Gene expression
  • NMR
  • Recombinant acylphosphatase
  • Synthetic gene

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics
  • Structural Biology

Cite this

Chemical synthesis and expression of a gene coding for human muscle acylphosphatase. / Modesti, Alessandra; Raugei, Giovanni; Taddei, Niccolo'; Marzocchini, Riccardo; Vecchi, Manuela; Camici, Guido; Manao, Giampaolo; Ramponi, Giampietro.

In: BBA - Gene Structure and Expression, Vol. 1216, No. 3, 14.12.1993, p. 369-374.

Research output: Contribution to journalArticle

Modesti, A, Raugei, G, Taddei, N, Marzocchini, R, Vecchi, M, Camici, G, Manao, G & Ramponi, G 1993, 'Chemical synthesis and expression of a gene coding for human muscle acylphosphatase', BBA - Gene Structure and Expression, vol. 1216, no. 3, pp. 369-374. https://doi.org/10.1016/0167-4781(93)90003-V
Modesti, Alessandra ; Raugei, Giovanni ; Taddei, Niccolo' ; Marzocchini, Riccardo ; Vecchi, Manuela ; Camici, Guido ; Manao, Giampaolo ; Ramponi, Giampietro. / Chemical synthesis and expression of a gene coding for human muscle acylphosphatase. In: BBA - Gene Structure and Expression. 1993 ; Vol. 1216, No. 3. pp. 369-374.
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