scholarly journals Characterization of the ToxB Gene from Pyrenophora tritici-repentis

2001 ◽  
Vol 14 (5) ◽  
pp. 675-677 ◽  
Author(s):  
J. Patrick Martinez ◽  
Sean A. Ottum ◽  
Shaukat Ali ◽  
Leonard J. Francl ◽  
Lynda M. Ciuffetti
Keyword(s):  
Amino Acid ◽  
Pichia Pastoris ◽  
Heterologous Expression ◽  
North Dakota ◽  
Signal Peptide ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Pyrenophora Tritici Repentis ◽  
Putative Signal Peptide

The ToxB gene was cloned and characterized from a race 5 isolate of Pyrenophora tritici-repentis from North Dakota. ToxB contains a 261-bp open reading frame that encodes a 23 amino acid putative signal peptide and a 64 amino acid host-selective toxin, Ptr ToxB. Analysis of Ptr ToxB from heterologous expression in Pichia pastoris confirms that ToxB encodes a host-selective toxin.

scholarly journals Characterization of a DivergentvanD-Type Resistance Element from the First Glycopeptide-Resistant Strain of Enterococcus faeciumIsolated in Brazil

2000 ◽  
Vol 44 (12) ◽  
pp. 3444-3446 ◽  
Author(s):  
Libera M. Dalla Costa ◽  
Peter E. Reynolds ◽  
Helena A. P. H. M. Souza ◽  
Dilair C. Souza ◽  
Marie-France I. Palepou ◽  
...  
Keyword(s):  
Amino Acid ◽  
Resistant Strain ◽  
Enterococcus Faecium ◽  
Amino Acid Identity ◽  
Open Reading Frame ◽  
Glycopeptide Resistance ◽  
Reading Frame ◽  
Acid Identity ◽  
Resistance Phenotype

ABSTRACT Enterococcus faecium 10/96A from Brazil was resistant to vancomycin (MIC, 256 μg/ml) but gave no amplification products with primers specific for known van genotypes. A 2,368-bp fragment of a van cluster contained one open reading frame encoding a peptide with 83% amino acid identity to VanHD, and a second encoding a d-alanine-d-lactate ligase with 83 to 85% identity to VanD. The divergent glycopeptide resistance phenotype was designated VanD4.

scholarly journals Heterologous, Expression, and Characterization of Thermostable Glucoamylase Derived fromAspergillus flavusNSH9 inPichia pastoris

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Kazi Muhammad Rezaul Karim ◽  
Ahmad Husaini ◽  
Md. Anowar Hossain ◽  
Ngieng Ngui Sing ◽  
Fazia Mohd Sinang ◽  
...  
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Pichia Pastoris ◽  
Aspergillus Flavus ◽  
Residual Activity ◽  
Open Reading Frame ◽  
Reading Frame ◽  
Higher Temperature ◽  
Positive Effect

A novel thermostable glucoamylase cDNA without starch binding domain (SBD) ofAspergillus flavusNSH9 was successfully identified, isolated, and overexpressed inPichia pastorisGS115. The complete open reading frame of glucoamylase fromAspergillus flavusNSH9 was identified by employing PCR that encodes 493 amino acids lacking in the SBD. The first 17 amino acids were presumed to be a signal peptide. The cDNA was cloned intoPichia pastorisand the highest expression of recombinant glucoamylase (rGA) was observed after 8 days of incubation period with 1% methanol. The molecular weight of the purified rGA was about 78 kDa and exhibited optimum catalytic activity at pH 5.0 and temperature of 70°C. The enzyme was stable at higher temperature with 50% of residual activity observed after 20 min at 90°C and 100°C. Low concentration of metal (Mg++, Fe++, Zn++, Cu++, and Pb++) had positive effect on rGA activity. This rGA has the potential for use and application in the saccharification steps, due to its thermostability, in the starch processing industries.

CHARACTERIZATION OF THE WHEAT cDNA ENCODING THE β SUBUNIT OF THE MITOCHONDRIAL ATP SYNTHASE

1996 ◽  
Vol 44 (2-3) ◽  
pp. 77-88 ◽  
Author(s):  
Suzy Abulafia ◽  
Dan Graur ◽  
Katrien Devos ◽  
Adina Breiman
Keyword(s):  
Amino Acids ◽  
Phylogenetic Analysis ◽  
Amino Acid ◽  
Open Reading Frame ◽  
Β Subunit ◽  
Reading Frame ◽  
Mitochondrial Atp Synthase ◽  
Group 1 Chromosomes ◽  
Group 1

A wheat cDNA encoding an open reading frame of 553 amino acids with a deduced amino acid sequence corresponding to the mitochondrial β subunit of the synthase was isolated. The expression of the β ATPase was investigated in leaves of 7-day-old wheat plants, and a decrease in the abundance of transcripts along the leaf was observed. The cDNA of the β ATPase was mapped on the group 1 chromosomes of wheat. Phylogenetic analysis of the mitochondrial β subunit of the ATPase complex is described.

scholarly journals Characterization of a 34-Kilodalton Protein ofMycobacterium leprae That Is Isologous to the Immunodominant 34-Kilodalton Antigen of Mycobacterium paratuberculosis

1998 ◽  
Vol 66 (11) ◽  
pp. 5576-5579 ◽  
Author(s):  
Fauzi S. Silbaq ◽  
Sang-Nae Cho ◽  
Stewart T. Cole ◽  
Patrick J. Brennan
Keyword(s):  
Sequence Analysis ◽  
Dna Sequence ◽  
Signal Peptide ◽  
Consensus Sequence ◽  
Mycobacterium Leprae ◽  
Open Reading Frame ◽  
Mycobacterium Paratuberculosis ◽  
Reading Frame ◽  
Serological Activity

ABSTRACT During DNA sequence analysis of cosmid L373 from theMycobacterium leprae genome, an open reading frame of 1.4 kb encoding a protein with some homology to the immunodominant 34-kDa protein of Mycobacterium paratuberculosis, but lacking significant serological activity, was detected. The DNA sequence predicted a signal peptide with a modified lipoprotein consensus sequence, but the protein proved to be devoid of lipid attachment.

scholarly journals Antigenic Characterization of ORF2 and ORF3 Proteins of Hepatitis E Virus (HEV)

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1385
Author(s):  
Giulia Pezzoni ◽  
Lidia Stercoli ◽  
Eleonora Pegoiani ◽  
Emiliana Brocchi
Keyword(s):  
Hepatitis E Virus ◽  
Hepatitis E ◽  
Recombinant Antigen ◽  
Open Reading Frame ◽  
Reading Frame ◽  
E Coli ◽  
Antigenic Characterization ◽  
Antigenic Properties ◽  
Open Reading Frame 2

To evaluate the antigenic properties of Hepatitis E Virus (HEV) Open Reading Frame 2 and 3 (ORF2 and ORF3) codified proteins, we expressed different portions of ORF2 and the entire ORF3 in E. coli, a truncated ORF2, was also expressed in baculovirus. A panel of 37 monoclonal antibodies (MAbs) was raised against ORF2 (1–660 amino acids) and MAbs were mapped and characterized using the ORF2 expressed portions. Selected HEV positive and negative swine sera were used to evaluate ORF2 and ORF3 antigens’ immunogenicity. The MAbs were clustered in six groups identifying six antigenic regions along the ORF2. Only MAbs binding to the sixth ORF2 antigenic region (394–608 aa) were found to compete with HEV positive sera and efficiently catch the recombinant antigen expressed in baculovirus. The ORF2 portion from 394–608 aa demonstrated to include most immunogenic epitopes with 85% of HEV positive swine sera reacting against the region from 461–544 aa. Only 5% of the selected HEV sera reacted against the ORF3 antigen.

scholarly journals Sequence and structure of mtr, an amino acid transport gene of Neurospora crassa

Genome ◽  
1991 ◽  
Vol 34 (4) ◽  
pp. 644-651 ◽  
Author(s):  
Kenneth Koo ◽  
W. Dorsey Stuart
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Neurospora Crassa ◽  
Rna Binding ◽  
Signal Sequence ◽  
Average Length ◽  
Open Reading Frame ◽  
Mrna Transcript ◽  
S1 Nuclease ◽  
Reading Frame

The gene product of the mtr locus of Neurospora crassa is required for the transport of neutral aliphatic and aromatic amino acids via the N system. We have previously cloned three cosmids containing Neurospora DNA that complement the mtr-6(r) mutant allele. The cloned DNAs were tightly linked to restriction fragment length polymorphisms that flank the mtr locus. A 2.9-kbp fragment from one cosmid was subcloned and found to complement the mtr-6(r) allele. Here we report the sequence of the fragment that hybridized to a poly(A)+ mRNA transcript of about 2300 nucleotides. We have identified an 845-bp open reading frame (ORF) having a 59-bp intron as the potential mtr ORF. S1 nuclease analysis of the transcript confirmed the transcript size and the presence of the intron. A second open reading frame was found upstream in the same reading frame as the mtr ORF and appears to be present in the mRNA transcript. The mtr ORF is predicted to encode a 261 amino acid polypeptide with a molecular mass of 28 613 Da. The proposed polypeptide exhibits six potential α-helical transmembrane domains with an average length of 23 amino acids, does not have a signal sequence, and contains amino acid sequence homologous to an RNA binding motif.Key words: sequence, membranes, ribonucleoprotein.

scholarly journals Gene Cloning and Molecular Characterization of a Two-Enzyme System Catalyzing the Oxidative Detoxification of β-Endosulfan

2002 ◽  
Vol 68 (12) ◽  
pp. 6237-6245 ◽  
Author(s):  
Tara D. Sutherland ◽  
Irene Horne ◽  
Robyn J. Russell ◽  
John G. Oakeshott
Keyword(s):  
Shuttle Vector ◽  
Open Reading Frame ◽  
Cosmid Library ◽  
Flavin Mononucleotide ◽  
Flavin Reductase ◽  
Reading Frame ◽  
Reductase Gene ◽  
Degradation Activity ◽  
Layer Chromatography

ABSTRACT The gram-positive bacterium Mycobacterium sp. strain ESD is able to use the cyclodiene insecticide endosulfan as a source of sulfur for growth. This activity is dependent on the absence of sulfite or sulfate in the growth medium. A cosmid library of strain ESD DNA was constructed in a Mycobacterium-Escherichia coli shuttle vector and screened for endosulfan-degrading activity in Mycobacterium smegmatis, a species that does not degrade endosulfan. Using this method, we identified a single cosmid that conferred sulfur-dependent endosulfan-degrading activity on the host strain. An open reading frame (esd) was identified within this cosmid that, when expressed behind a constitutive promoter in a mycobacterial expression vector, conferred sulfite- and sulfate-independent β-endosulfan degradation activity on the recombinant strain. The translation product of this gene (Esd) had up to 50% sequence identity with an unusual family of monooxygenase enzymes that use reduced flavins, provided by a separate flavin reductase enzyme, as cosubstrates. An additional partial open reading frame was located upstream of the Esd gene that had sequence homology to the same monooxygenase family. A flavin reductase gene, identified in the M. smegmatis genome, was cloned, expressed, and used to provide reduced flavin mononucleotide for Esd in enzyme assays. Thin-layer chromatography and gas chromatography analyses of the enzyme assay mixtures revealed the disappearance of β-endosulfan and the appearance of the endosulfan metabolites, endosulfan monoaldehyde and endosulfan hydroxyether. This suggests that Esd catalyzes the oxygenation of β-endosulfan to endosulfan monoaldehyde and endosulfan hydroxyether. Esd did not degrade either α-endosulfan or the metabolite of endosulfan, endosulfan sulfate.

scholarly journals Cloning and characterization of α-pinene synthase from Chamaecyparis formosensis Matsum

Holzforschung ◽  
2009 ◽  
Vol 63 (1) ◽  
Author(s):  
Fang-Hua Chu ◽  
Pei-Min Kuo ◽  
Yu-Rong Chen ◽  
Sheng-Yang Wang
Keyword(s):  
Major Product ◽  
Open Reading Frame ◽  
Terpene Synthase ◽  
Geranyl Diphosphate ◽  
Reading Frame ◽  
E Coli ◽  
Phylogenetic Taxonomy ◽  
Chamaecyparis Formosensis ◽  
Pcr Method

AbstractAnalyzing the gene sequences of terpene synthase (TPS) may contribute to a better understanding of terpenes biosynthesis and evolution of phylogenetic taxonomy.Chamaecyparis formosensisis an endemic and precious conifer of Taiwan. To understand the biosynthesis mechanism of terpenes in this tree, a full length of putative mono-TPS, named asCf-Pin(GeneBank accession no. EU099434), was obtained by PCR method and RACE extension. TheCf-Pinhas an 1887-bp open reading frame and encodes 628 amino acids. To identify the function ofCf-Pin,the recombinant protein fromEscherichia coliwas incubated with geranyl diphosphate, produced one major product, the structure of which was elucidated. GC/MS analysis and matching of retention time and mass spectrum with authentic standards revealed that this product isα-pinene. This is the first report of cloning of a mono-TPS and functionally expressed inE. coliand which could be identified asα-pinene synthase from a Cupressaceae conifer.

Heterologous expression and characterization of l-amino acid deaminases from Proteus mirabilis in Escherichia coli

2008 ◽  
Vol 136 ◽  
pp. S300 ◽  
Author(s):  
Jin-Oh Baek ◽  
Jeong-Woo Seo ◽  
Ohsuk Kwon ◽  
Su-Il Seong ◽  
Ik-Hwan Kim ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acid ◽  
Heterologous Expression ◽  
Proteus Mirabilis
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