Journal:

Article Title: Identification of two epoxide hydrolases in Caenorhabditis elegans that metabolize mammalian lipid signaling molecules

doi: 10.1016/j.abb.2008.01.016

Figure Lengend Snippet: Protein BLAST results

Article Snippet: Cloning of the hydrolases A tBLASTx search of the NCBI genomic database of C. elegans employing Gallus gallus (Genbank accession number Q120010) and Xenopus tropicalis (Genbank accession number BC078066) sEH nucleotide and translated amino acid sequences returned five soluble epoxide hydrolase hits.

Techniques:

Journal:

Article Title: Identification of two epoxide hydrolases in Caenorhabditis elegans that metabolize mammalian lipid signaling molecules

doi: 10.1016/j.abb.2008.01.016

Figure Lengend Snippet: Alignment of the vertebrate sEH homologs with the nematode enzymes and bacterial HLD. The mammalian epoxide hydrolase “catalytic triad” residues are marked by closed triangles. The tyrosines thought to polarize the epoxide moiety of the epoxide hydrolase substrate are marked by open squares. HSEH = sEH from H. sapiens (Genbank accession no. L05779); GGSEH = sEH from G. gallus (DQ120010); XTSEH = sEH from X. tropicalis (BC075370); XAHLD = HLD from X. autotrophicus (M26950).

Article Snippet: Cloning of the hydrolases A tBLASTx search of the NCBI genomic database of C. elegans employing Gallus gallus (Genbank accession number Q120010) and Xenopus tropicalis (Genbank accession number BC078066) sEH nucleotide and translated amino acid sequences returned five soluble epoxide hydrolase hits.

Techniques:

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: SDS-PAGE demonstrating the expression level and solubility of p30 (a) and SUMO-p30 (b) in Escherichia coli after IPTG induction. M: Molecular weight marker; 1: No IPTG induction of protein expression; 2: IPTG induced protein expression; 3: Protein in the soluble fraction; 4: Protein in the insoluble fraction. IPTG=Isopropyl β-D-thiogalactoside. SUMO=Small ubiquitin-like modifier, SDS-PAGE=Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: SDS Page, Expressing, Solubility, Molecular Weight, Marker, Ubiquitin Proteomics, Polyacrylamide Gel Electrophoresis

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: SDS-PAGE showing protein profiles of purified p30 and SUMO-p30. Qualities of p30 (a, b, and c) and SUMO-p30 (d, e, and f) partially purified by ammonium sulphate (AS) precipitation (b and e) followed by Ni-IDA affinity chromatography (c and f) can be observed to be higher than those purified with affinity chromatography alone (a and d). M: molecular weight marker. B: Before AS precipitation. A: After AS precipitation. SUMO=Small ubiquitin-like modifier, SDS-PAGE=Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, IDA=Iminodiacetic acid.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: SDS Page, Purification, Affinity Chromatography, Molecular Weight, Marker, Ubiquitin Proteomics, Polyacrylamide Gel Electrophoresis

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: Percentage optimization of saturated ammonium sulfate suitable for precipitating p30 and SUMO-p30.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques:

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: SDS-PAGE showing the results of enzymatic cleavage and removal of SUMO tag. SUMO-p30 cleaved with SUMO protease 1 (a). Removal of 6×His-SUMO tag, accomplished by reintroducing the protein solution into the Ni-IDA resin column (b). M: Molecular weight marker; Uncut: SUMO-p30; Cut: SUMO-cleaved p30; B: before removing 6ÍHis-SUMO tag, FT: SUMO-cleaved p30 in the flow-through fraction, W1-3: Washed fractions containing 20-, 50-, and 100-mM imidazole, respectively. E: Eluted fraction containing 250 mM imidazole. SUMO=Small ubiquitin-like modifier, SDS-PAGE=Sodium dodecyl sulfate-polyacrylamide gel electrophoresis, IDA=Iminodiacetic acid.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: SDS Page, Molecular Weight, Marker, Ubiquitin Proteomics, Polyacrylamide Gel Electrophoresis

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: Western blots demonstrating the immunogenicity of p30 proteins. 5 μL of each protein loaded into the wells of polyacrylamide gels for electrophoresis. The gels are blotted onto nitrocellulose membranes before incubating with a monoclonal antibody specific to p30 (a) or a swine convalescent serum (b). All versions of p30 can be observed to react with a monoclonal antibody specific to p30 (a) and a swine convalescent serum (b), while the capsid protein of porcine circovirus type 2 (a) and glycoprotein G of bovine ephemeral fever virus (b) can be observed not to react. M: Molecular weight marker.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: Western Blot, Immunopeptidomics, Electrophoresis, Virus, Molecular Weight, Marker

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: Comparison of p30 and SUMO-p30 with and without partial purification by ammonium sulfate precipitation. The images show protein profiles in SDS-PAGE (a), their immunogenicity in western blots (b), and the protein yields in milligrams per liter of Escherichia coli culture (c). M: Molecular weight marker. AS: Ammonium sulfate precipitation. SUMO=Small ubiquitin-like modifier, SDS-PAGE=Sodium dodecyl sulfate-polyacrylamide gel electrophoresis.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: Comparison, Purification, SDS Page, Immunopeptidomics, Western Blot, Molecular Weight, Marker, Ubiquitin Proteomics, Polyacrylamide Gel Electrophoresis

Journal: Veterinary World

Article Title: Small ubiquitin-like modifier-tag and modified protein purification significantly increase the quality and quantity of recombinant African swine fever virus p30 protein

doi: 10.14202/vetworld.2024.1157-1167

Figure Lengend Snippet: Performance of indirect ELISA based on SUMO-cleaved p30. The cutoff value of the p30 indirect ELISA (a) was 0.291 by setting at mean OD of negative sera plus three-fold standard deviations (X̅ + 3SD). Scatter plots of OD450 of swine serum samples obtained from the p30 indirect ELISA showing a significant difference in the OD450 of positive and negative serum samples (b). The p30 indirect ELISA does not react with serums raised against the porcine reproductive and respiratory syndrome virus, swine influenza virus, and porcine circovirus type 2 (c). SUMO=Small ubiquitin-like modifier, ELISA=Enzyme-linked immunosorbent assay, OD=Optical density.

Article Snippet: The CP204L gene encoding p30 protein (GenBank accession number: OR567419) cloned in pUC57 (Genescript, Piscataway, NJ, USA) was used as a template to amplify the CP204L gene with a pair of specific primers: forward primer; 5’TAGCATGCATGGCATCAGGAGGAGC- 3’ (containing a Sph I restriction site) and reverse primer; 5’-TAGTCGACTTAGGTACTGTAACGCAG -3’ (containing a Sal I restriction site) using PhusionTM high-fidelity DNA polymerase (Thermo Fisher Scientific, Waltham, Massachusetts, USA).

Techniques: Indirect ELISA, Virus, Ubiquitin Proteomics, Enzyme-linked Immunosorbent Assay

Journal: Journal of Biomedical Science

Article Title: Structural and functional characterization of MERS coronavirus papain-like protease

doi: 10.1186/1423-0127-21-54

Figure Lengend Snippet: The kinetic parameters and DUB activity of MERS - CoV PL pro

Article Snippet: The sequence of MERS-CoV PL pro (GenBank accession number NC_019843.2; polyprotein residues 1484–1800) was synthesized (MDBio Inc.), digested by Nco I- Xho I and then inserted into the pET-28a(+) vector (Novagen).

Techniques: Activity Assay, Mutagenesis