Synthesis, cloning and expression of a synthetic gene for high potential iron protein from Chromatium vinosum.
Identifieur interne : 002942 ( Ncbi/Merge ); précédent : 002941; suivant : 002943Synthesis, cloning and expression of a synthetic gene for high potential iron protein from Chromatium vinosum.
Auteurs : A. Agarwal ; J. Tan ; M. Eren ; A. Tevelev ; S M Lui ; J A CowanSource :
- Biochemical and biophysical research communications [ 0006-291X ] ; 1993.
Descripteurs français
- KwdFr :
- Cartographie de restriction, Chromatium (génétique), Chromatium (métabolisme), Clonage moléculaire, Complexe protéique du centre réactionnel de la photosynthèse, Données de séquences moléculaires, Expression des gènes, Ferrosulfoprotéines (), Ferrosulfoprotéines (biosynthèse), Ferrosulfoprotéines (génétique), Ferrédoxines (biosynthèse), Gènes de synthèse, Oligodésoxyribonucléotides (synthèse chimique), Protéines bactériennes (biosynthèse), Protéines recombinantes (), Protéines recombinantes (biosynthèse), Spectroscopie par résonance magnétique, Séquence d'acides aminés, Séquence nucléotidique.
- MESH :
- biosynthèse : Ferrosulfoprotéines, Ferrédoxines, Protéines bactériennes, Protéines recombinantes.
- génétique : Chromatium, Ferrosulfoprotéines.
- métabolisme : Chromatium.
- synthèse chimique : Oligodésoxyribonucléotides.
- Cartographie de restriction, Clonage moléculaire, Complexe protéique du centre réactionnel de la photosynthèse, Données de séquences moléculaires, Expression des gènes, Ferrosulfoprotéines, Gènes de synthèse, Protéines recombinantes, Spectroscopie par résonance magnétique, Séquence d'acides aminés, Séquence nucléotidique.
English descriptors
- KwdEn :
- Amino Acid Sequence, Bacterial Proteins (biosynthesis), Base Sequence, Chromatium (genetics), Chromatium (metabolism), Cloning, Molecular, Ferredoxins (biosynthesis), Gene Expression, Genes, Synthetic, Iron-Sulfur Proteins (biosynthesis), Iron-Sulfur Proteins (chemistry), Iron-Sulfur Proteins (genetics), Magnetic Resonance Spectroscopy, Molecular Sequence Data, Oligodeoxyribonucleotides (chemical synthesis), Photosynthetic Reaction Center Complex Proteins, Recombinant Proteins (biosynthesis), Recombinant Proteins (chemistry), Restriction Mapping.
- MESH :
- chemical , biosynthesis : Bacterial Proteins, Ferredoxins, Iron-Sulfur Proteins, Recombinant Proteins.
- chemical , chemical synthesis : Oligodeoxyribonucleotides.
- chemical , chemistry : Iron-Sulfur Proteins, Recombinant Proteins.
- genetics : Chromatium, Iron-Sulfur Proteins.
- metabolism : Chromatium.
- Amino Acid Sequence, Base Sequence, Cloning, Molecular, Gene Expression, Genes, Synthetic, Magnetic Resonance Spectroscopy, Molecular Sequence Data, Photosynthetic Reaction Center Complex Proteins, Restriction Mapping.
Abstract
A synthetic gene encoding the peptide sequence for the low molecular weight (M(r) approximately 9600 Da) high-potential iron protein (HiPIP) from the photosynthetic bacterium Chromatium vinosum has been constructed by shotgun ligation of twelve complimentary oligonucleotides varying in size from 42-mers to 48-mers. After cloning the gene into a pET-21d(+) vector, expression of holoprotein in yields of 35 mg/liter of culture was obtained following induction with isopropyl-beta-D-thiogalactoside (IPTG). The recombinant protein was characterized by electronic absorption, 1H NMR, electrochemistry, N-terminal sequencing and amino acid analysis. This is the first example of the expression of a high potential ferredoxin containing a fully constituted [Fe4S4] cluster.
DOI: 10.1006/bbrc.1993.2626
PubMed: 7916611
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pubmed:7916611Le document en format XML
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After cloning the gene into a pET-21d(+) vector, expression of holoprotein in yields of 35 mg/liter of culture was obtained following induction with isopropyl-beta-D-thiogalactoside (IPTG). The recombinant protein was characterized by electronic absorption, 1H NMR, electrochemistry, N-terminal sequencing and amino acid analysis. 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Agarwal</name><name sortKey="Cowan, J A" sort="Cowan, J A" uniqKey="Cowan J" first="J A" last="Cowan">J A Cowan</name><name sortKey="Eren, M" sort="Eren, M" uniqKey="Eren M" first="M" last="Eren">M. Eren</name><name sortKey="Lui, S M" sort="Lui, S M" uniqKey="Lui S" first="S M" last="Lui">S M Lui</name><name sortKey="Tan, J" sort="Tan, J" uniqKey="Tan J" first="J" last="Tan">J. Tan</name><name sortKey="Tevelev, A" sort="Tevelev, A" uniqKey="Tevelev A" first="A" last="Tevelev">A. Tevelev</name></noCountry></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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