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

doi:10.1016/0167-4781(93)90003-V

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

Istituto Europeo di Oncologia

Research output: Contribution to journal › Article › peer-review

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 language	English
Pages (from-to)	369-374
Number of pages	6
Journal	BBA - Gene Structure and Expression
Volume	1216
Issue number	3
DOIs	https://doi.org/10.1016/0167-4781(93)90003-V
Publication status	Published - Dec 14 1993

Keywords

(Human)
Gene expression
NMR
Recombinant acylphosphatase
Synthetic gene

ASJC Scopus subject areas

Biochemistry
Biophysics
Genetics
Structural Biology

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10.1016/0167-4781(93)90003-V

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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 journal › Article › peer-review

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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.",

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AU - Taddei, Niccolo'

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AU - Vecchi, Manuela

AU - Camici, Guido

AU - Manao, Giampaolo

AU - Ramponi, Giampietro

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

Abstract

Keywords

ASJC Scopus subject areas

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