Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: SDS-DNA PAGE of IF cells of H. obtusa. Calf thymus DNA was used. ( a ) CBB stained gel; ( b ) immunoblot with mAb3D1B9C4 specific for PRP1; ( c ) EB stained gel. Arrows, PRP1. Arrowheads, other EB-staining negative bands showing positions of DNA-protein complexes (see ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: IF-cells, Staining, Western Blot

Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: SDS-PAGE and immunoblots of fractions of DNA affinity column chromatography. Calf thymus DNA was used. ( a ) Fractions 1–10 eluted with Na, K-PB; ( b ) fractions 1′–10′ eluted with Na, K-PB containing 0.2 M NaCl. Upper, silver-stained gel. Lower, immunoblot with mAb3D1B9C4. ( c ) High magnification of a fraction number 3′ using silver staining. Not only PRP1 but also several silver staining bands were also confirmed. ( d ) CBB stained gel of IF cells of H. obtusa . Arrow, PRP1. Arrowheads, other bands eluted. Bands in ( c ) match the bands in ( d ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: SDS Page, Western Blot, Affinity Column, Chromatography, Staining, Silver Staining, IF-cells

Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: SDS-DNA PAGE of IF cells of H. obtusa . P. caudatum DNA was used. ( a ) CBB stained gel. ( b ) Immunoblot with mAb3D1B9C4 specific for PRP1. ( c ) EB stained gel. Arrows, 63-kDa PRP1. It should be noted that not only PRP1 but also many EB-negative bands that show the positions of DNA-protein complexes appeared (see ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: IF-cells, Staining, Western Blot

Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: DNA affinity chromatography of H. obtusa IFs. P. caudatum DNA was used. ( a ) Silver stained SDS-PAGE gel; ( b ) immunoblotting with the monoclonal antibody mAb3D1B9C4 specific for the 63-kDa periplasmic protein PRP1 of H. obtusa . Lane 1, supernatant of sonicated H. obtusa before applying to DNA affinity chromatography column; lane 2, first elute of supernatant of sonicated H. obtusa after applying to the column; lane 3, first elute by elution buffer containing 0.02 M NaCl; lane 4, first eluate by 0.1 M NaCl elution buffer; lane 5, first eluate by 0.2 M NaCl elution buffer; lane 6, first eluate by 0.3 M NaCl elution buffer; lane 7, first eluate by 2 M NaCl elution buffer; arrows, PRP1. Eluates from each column were collected in every 200 μL, and the first eluate from each column was used for SDS-PAGE and immunoblot with mAb. Note that not only PRP1 but also several silver stained bands were confirmed in lane 4 of ( a ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: Affinity Chromatography, Staining, SDS Page, Western Blot, Sonication, Affinity Column

Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: SDS-DNA PAGE of IF cells of H. obtusa . P. caudatum DNA was used. ( a ) CBB stained gel. ( b ) Immunoblot with mAb3D1B9C4 specific for PRP1. ( c ) EB stained gel. Arrows, 63-kDa PRP1. It should be noted that not only PRP1 but also many EB-negative bands that show the positions of DNA-protein complexes appeared (see ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: IF-cells, Staining, Western Blot

Journal: Microorganisms

Article Title: A 63-kDa Periplasmic Protein of the Endonuclear Symbiotic Bacterium Holospora obtusa Secreted to the Outside of the Bacterium during the Early Infection Process Binds Weakly to the Macronuclear DNA of the Host Paramecium caudatum

doi: 10.3390/microorganisms11010155

Figure Lengend Snippet: DNA affinity chromatography of H. obtusa IFs. P. caudatum DNA was used. ( a ) Silver stained SDS-PAGE gel; ( b ) immunoblotting with the monoclonal antibody mAb3D1B9C4 specific for the 63-kDa periplasmic protein PRP1 of H. obtusa . Lane 1, supernatant of sonicated H. obtusa before applying to DNA affinity chromatography column; lane 2, first elute of supernatant of sonicated H. obtusa after applying to the column; lane 3, first elute by elution buffer containing 0.02 M NaCl; lane 4, first eluate by 0.1 M NaCl elution buffer; lane 5, first eluate by 0.2 M NaCl elution buffer; lane 6, first eluate by 0.3 M NaCl elution buffer; lane 7, first eluate by 2 M NaCl elution buffer; arrows, PRP1. Eluates from each column were collected in every 200 μL, and the first eluate from each column was used for SDS-PAGE and immunoblot with mAb. Note that not only PRP1 but also several silver stained bands were confirmed in lane 4 of ( a ).

Article Snippet: For SDS–DNA PAGE, a 10% ( w / v ) separation gel containing 10 μg mL –1 calf thymus DNA (Worthington Biochemical Corp., Lakewood, NJ, USA) or 10 μg mL –1 P. caudatum whole genome DNA was used.

Techniques: Affinity Chromatography, Staining, SDS Page, Western Blot, Sonication, Affinity Column