vector phy304 Search Results


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  • 99
    New England Biolabs gibson assembly cloning kit
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    Fisher Bioreagents b anthracis
    Heteromultimerization by PCVRs Lysates from B. <t>anthracis</t> atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible AtxA-His (pUTE991), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AtxA, AcpA, and GFP.
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    Heteromultimerization by PCVRs Lysates from B. <t>anthracis</t> atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible AtxA-His (pUTE991), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AtxA, AcpA, and GFP.
    Miniprep, supplied by IBI Scientific, used in various techniques. Bioz Stars score: 92/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Heteromultimerization by PCVRs Lysates from B. anthracis atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible AtxA-His (pUTE991), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AtxA, AcpA, and GFP.

    Journal: Molecular microbiology

    Article Title: Regulons and Protein-Protein Interactions of PRD-containing Bacillus anthracis Virulence Regulators Reveal Overlapping but Distinct Functions

    doi: 10.1111/mmi.13961

    Figure Lengend Snippet: Heteromultimerization by PCVRs Lysates from B. anthracis atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible AtxA-His (pUTE991), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AtxA, AcpA, and GFP.

    Article Snippet: B. anthracis containing the pHY304-derived construct was cultivated at 41°C (the non-permissive replication temperature) in medium supplemented with erythromycin to select for isolates in which the plasmid incorporated into one of the acpA flanking regions via homologous recombination.

    Techniques: Incubation, Affinity Purification, Purification, SDS Page, Western Blot

    Homomultimerization AcpA and AcpB Lysates from B. anthracis atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible (A) AcpA-His (pUTE1090), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); (B) AcpB-His (pUTE1091), AcpB-FLAG (pUTE1093), or GFP-FLAG (pUTE1013) were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AcpA-His, AcpA-FLAG, AcpB-His, AcpB-FLAG, and GFP-FLAG; (C) FLAG-tagged AcpA (pUTE1079) and AcpB (pUTE1093) were induced by IPTG in a B. anthracis atxA- null pXO1+ pXO2- strain. Lysates were incubated with or without the crosslinking agent BMH and subjected to SDS-PAGE and Western blot. Proteins were detected with α-FLAG antibody. (D) Affinity purified AcpA-His from B. anthracis ANR-1 (pUTE1090) incubated with or without BMH and subjected to SDS-PAGE and Western blot. Proteins were detected with α-His antibody.

    Journal: Molecular microbiology

    Article Title: Regulons and Protein-Protein Interactions of PRD-containing Bacillus anthracis Virulence Regulators Reveal Overlapping but Distinct Functions

    doi: 10.1111/mmi.13961

    Figure Lengend Snippet: Homomultimerization AcpA and AcpB Lysates from B. anthracis atxA- null pXO1+ pXO2- strains (UT423) containing plasmids that encode IPTG-inducible (A) AcpA-His (pUTE1090), AcpA-FLAG (pUTE1079), or GFP-FLAG (pUTE1013); (B) AcpB-His (pUTE1091), AcpB-FLAG (pUTE1093), or GFP-FLAG (pUTE1013) were co-incubated as indicated, then co-affinity purified with Ni 2+ -NTA resin. Proteins present in the mixed lysates prior to (Load, lanes 1–3) and after purification (Eluate, lanes 4–6) were subjected to SDS-PAGE and Western blot with α-His and α-FLAG antibodies as indicated. Arrows indicate the predicted sizes of AcpA-His, AcpA-FLAG, AcpB-His, AcpB-FLAG, and GFP-FLAG; (C) FLAG-tagged AcpA (pUTE1079) and AcpB (pUTE1093) were induced by IPTG in a B. anthracis atxA- null pXO1+ pXO2- strain. Lysates were incubated with or without the crosslinking agent BMH and subjected to SDS-PAGE and Western blot. Proteins were detected with α-FLAG antibody. (D) Affinity purified AcpA-His from B. anthracis ANR-1 (pUTE1090) incubated with or without BMH and subjected to SDS-PAGE and Western blot. Proteins were detected with α-His antibody.

    Article Snippet: B. anthracis containing the pHY304-derived construct was cultivated at 41°C (the non-permissive replication temperature) in medium supplemented with erythromycin to select for isolates in which the plasmid incorporated into one of the acpA flanking regions via homologous recombination.

    Techniques: Incubation, Affinity Purification, Purification, SDS Page, Western Blot

    Edema Factor Production by individual PCVRs Expression of recombinant atxA, acpA, and acpB was induced with IPTG to yield either native or overexpressed steady state protein levels (Native - 5 μM, 5 μM, 100 μM respectively; Overexpressed - 50 μM, 50 μM, 500 μM respectively) during growth in CA medium supplemented with dissolved bicarbonate in 5% CO 2 atmosphere. Samples of culture supernates were subjected to slot blot Western analysis using rabbit anti-EF serum raised against B. anthracis edema factor.

    Journal: Molecular microbiology

    Article Title: Regulons and Protein-Protein Interactions of PRD-containing Bacillus anthracis Virulence Regulators Reveal Overlapping but Distinct Functions

    doi: 10.1111/mmi.13961

    Figure Lengend Snippet: Edema Factor Production by individual PCVRs Expression of recombinant atxA, acpA, and acpB was induced with IPTG to yield either native or overexpressed steady state protein levels (Native - 5 μM, 5 μM, 100 μM respectively; Overexpressed - 50 μM, 50 μM, 500 μM respectively) during growth in CA medium supplemented with dissolved bicarbonate in 5% CO 2 atmosphere. Samples of culture supernates were subjected to slot blot Western analysis using rabbit anti-EF serum raised against B. anthracis edema factor.

    Article Snippet: B. anthracis containing the pHY304-derived construct was cultivated at 41°C (the non-permissive replication temperature) in medium supplemented with erythromycin to select for isolates in which the plasmid incorporated into one of the acpA flanking regions via homologous recombination.

    Techniques: Expressing, Recombinant, Dot Blot, Western Blot

    Activity and multimerization of AcpA and AcpB EIIB-like domain truncation mutants UT423 strains expressing AcpA-ΔEIIB-His (pUTE1125), AcpA-FLAG (pUTE1079), AcpB-ΔEIIB-His (pUTE1126), or AcpB-FLAG (pUTE1093) were cultured in CA medium supplemented with dissolved bicarbonate in 5% CO 2 atmosphere and induced with 30–50 μM IPTG. A. Cell lysates containing IPTG-induced proteins were treated with crosslinking agent BMH or vehicle alone (DMSO). Molecular weights of protein standards are listed. B. The β-galactosidase activity of B. anthracis strains harboring the P capB-lacZ ).. Errors represent ±1 SD.

    Journal: Molecular microbiology

    Article Title: Regulons and Protein-Protein Interactions of PRD-containing Bacillus anthracis Virulence Regulators Reveal Overlapping but Distinct Functions

    doi: 10.1111/mmi.13961

    Figure Lengend Snippet: Activity and multimerization of AcpA and AcpB EIIB-like domain truncation mutants UT423 strains expressing AcpA-ΔEIIB-His (pUTE1125), AcpA-FLAG (pUTE1079), AcpB-ΔEIIB-His (pUTE1126), or AcpB-FLAG (pUTE1093) were cultured in CA medium supplemented with dissolved bicarbonate in 5% CO 2 atmosphere and induced with 30–50 μM IPTG. A. Cell lysates containing IPTG-induced proteins were treated with crosslinking agent BMH or vehicle alone (DMSO). Molecular weights of protein standards are listed. B. The β-galactosidase activity of B. anthracis strains harboring the P capB-lacZ ).. Errors represent ±1 SD.

    Article Snippet: B. anthracis containing the pHY304-derived construct was cultivated at 41°C (the non-permissive replication temperature) in medium supplemented with erythromycin to select for isolates in which the plasmid incorporated into one of the acpA flanking regions via homologous recombination.

    Techniques: Activity Assay, Expressing, Cell Culture