Construction of synthetic open reading frame encoding human interferon alpha 2b for high expression in Escherichia coli and characterization of its gene product

ABSTRACT The open reading frame at 86.7 min on the Escherichia coli chromosome, “yigC,” complemented aubiD mutant strain, AN66, indicating that yigCis the ubiD gene. The gene product, a 497-amino-acid-residue protein, showed extensive homology to the UPF 00096 family of proteins in the Swiss-Prot database.

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Characterization of the tol-pal region of Escherichia coli K-12: translational control of tolR expression by TolQ and identification of a new open reading frame downstream of pal encoding a periplasmic protein.

Journal of Bacteriology ◽

10.1128/jb.178.14.4031-4038.1996 ◽

1996 ◽

Vol 178 (14) ◽

pp. 4031-4038 ◽

Cited By ~ 55

Author(s):

A Vianney ◽

M M Muller ◽

T Clavel ◽

J C Lazzaroni ◽

R Portalier ◽

...

Keyword(s):

Escherichia Coli ◽

Translational Control ◽

Open Reading Frame ◽

Periplasmic Protein ◽

Reading Frame ◽

K 12

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3-hydroxy-3-methylglutaryl coenzyme A reductase of Sulfolobus solfataricus: DNA sequence, phylogeny, expression in Escherichia coli of the hmgA gene, and purification and kinetic characterization of the gene product.

Journal of Bacteriology ◽

10.1128/jb.179.11.3632-3638.1997 ◽

1997 ◽

Vol 179 (11) ◽

pp. 3632-3638 ◽

Cited By ~ 32

Author(s):

D A Bochar ◽

J R Brown ◽

W F Doolittle ◽

H P Klenk ◽

W Lam ◽

...

Keyword(s):

Escherichia Coli ◽

Gene Product ◽

Dna Sequence ◽

Coenzyme A ◽

Sulfolobus Solfataricus ◽

Kinetic Characterization ◽

Expression In Escherichia Coli ◽

Coenzyme A Reductase

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Functional Characterization of Alanine Racemase fromSchizosaccharomyces pombe: a Eucaryotic Counterpart to Bacterial Alanine Racemase

Journal of Bacteriology ◽

10.1128/jb.183.7.2226-2233.2001 ◽

2001 ◽

Vol 183 (7) ◽

pp. 2226-2233 ◽

Cited By ~ 53

Author(s):

Takuma Uo ◽

Tohru Yoshimura ◽

Naotaka Tanaka ◽

Kaoru Takegawa ◽

Nobuyoshi Esaki

Keyword(s):

Escherichia Coli ◽

Nitrogen Source ◽

Functional Characterization ◽

Open Reading Frame ◽

Alanine Racemase ◽

Sole Nitrogen Source ◽

Recombinant Yeast ◽

Reading Frame ◽

Putative Protein

ABSTRACT Schizosaccharomyces pombe has an open reading frame, which we named alr1 +, encoding a putative protein similar to bacterial alanine racemase. We cloned thealr1 + gene in Escherichia coli and purified the gene product (Alr1p), with an M rof 41,590, to homogeneity. Alr1p contains pyridoxal 5′-phosphate as a coenzyme and catalyzes the racemization of alanine with apparentKm and V max values as follows: for l-alanine, 5.0 mM and 670 μmol/min/mg, respectively, and for d-alanine, 2.4 mM and 350 μmol/min/mg, respectively. The enzyme is almost specific to alanine, but l-serine and l-2-aminobutyrate are racemized slowly at rates 3.7 and 0.37% of that ofl-alanine, respectively. S. pombe usesd-alanine as a sole nitrogen source, but deletion of thealr1 + gene resulted in retarded growth on the same medium. This indicates that S. pombe has catabolic pathways for both enantiomers of alanine and that the pathway forl-alanine coupled with racemization plays a major role in the catabolism of d-alanine. Saccharomyces cerevisiae differs markedly from S. pombe: S. cerevisiae uses l-alanine but notd-alanine as a sole nitrogen source. Moreover,d-alanine is toxic to S. cerevisiae. However, heterologous expression of the alr1 + gene enabled S. cerevisiae to grow efficiently ond-alanine as a sole nitrogen source. The recombinant yeast was relieved from the toxicity of d-alanine.

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Cloning and characterization of theddsAgene encoding decaprenyl diphosphate synthase fromRhodobacter capsulatusB10

Canadian Journal of Microbiology ◽

10.1139/w06-080 ◽

2006 ◽

Vol 52 (12) ◽

pp. 1141-1147 ◽

Cited By ~ 2

Author(s):

Xinyi Liu ◽

Haizhen Wu ◽

Jiang Ye ◽

Qinsheng Yuan ◽

Huizhan Zhang

Keyword(s):

Gene Expression ◽

Escherichia Coli ◽

Rhodobacter Capsulatus ◽

Open Reading Frame ◽

High Similarity ◽

Reading Frame ◽

Endogenous Production ◽

Genome Library

A decaprenyl diphosphate synthase gene (ddsA, GenBank accession No. DQ191802) was cloned from Rhodobacter capsulatus B10 by constructing and screening the genome library. An open reading frame of 1002 bp was revealed from sequence analysis. The deduced polypeptide consisted of 333 amino acids residues with an molecular mass of about 37 kDa. The DdsA protein contained the conserved amino acid sequence (DDXXD) of E-type polyprenyl diphosphate synthase and showed high similarity to others. In contrast, DdsA showed only 39% identity to a solanesyl diphosphate synthase cloned from R. capsulatus SB1003. DdsA was expressed successfully in Escherichia coli. Assaying the enzyme in vivo found it made E.coli synthesize UQ-10 in addition to the endogenous production UQ-8.Key words: ubiquinone, polyprenyl diphosphate synthase, gene expression, Rhodobacter capsulatus.

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