puc19 Takara Search Results


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  • 79
    TaKaRa plasmid puc19 clontech takara
    Plasmid Puc19 Clontech Takara, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    TaKaRa puc 19
    QdNO-induced DNA strand breakage. (A, B) E . coli DH5α cells harboring the pBR322 plasmid were incubated with the indicated concentration of CYA (A) or OLA (B) at 37°C for 2 h. TPZ was used as the positive control drug. (C) Inhibition of the degradation of <t>pUC-19</t> plasmid in E . coli DH5α by free radical scavengers. Bacteria harboring pUC-19 plasmid were incubated with 100 mM mannitol, thiourea, acetone, β-ME, ethanol, methanol and NaN 3 (from lane 2 to lane 8), and after 4 μg/ml CYA was added, the cells were incubated at 37°C for 2 h. The untreated bacteria (lane 1) and the bacteria treated only with CYA but no scavengers (lane 9) were used as the blank and the control, respectively. (D, E) Supercoiled pBR322 DNA (10 μg/ml) was incubated with the indicated concentration of CYA (D) or OLA (E) in the presence of xanthine oxidase/xanthine at 37°C for 0.5 h. The plasmids isolated from the treated bacteria (A-C) or the treated plasmids (D, E) were electrophoretically separated. XO/X, xanthine oxidase/xanthine; LM, linear DNA marker; SM, supercoiled DNA marker. SC, L and OC indicate supercoiled, linear and open circular DNA, respectively.
    Puc 19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa plasmid puc 19
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Plasmid Puc 19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 82 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    TaKaRa bamhi digested puc19
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Bamhi Digested Puc19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 89/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa puc19 derivative pegfp
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Puc19 Derivative Pegfp, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    TaKaRa ndei ecori digested plasmid puc19
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Ndei Ecori Digested Plasmid Puc19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    TaKaRa puc19 suicide vector
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Puc19 Suicide Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 81/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    TaKaRa puc19 t vector system
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Puc19 T Vector System, supplied by TaKaRa, used in various techniques. Bioz Stars score: 78/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    TaKaRa ecori digested puc19 dna
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Ecori Digested Puc19 Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 82/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa puc19 derivative ppd16 43
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Puc19 Derivative Ppd16 43, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa sali smai digested puc19
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Sali Smai Digested Puc19, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    TaKaRa xhoi digested puc19 mcp dna
    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from <t>pUC19</t> were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.
    Xhoi Digested Puc19 Mcp Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 80/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    TaKaRa puc18 dna
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Puc18 Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 77/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    TaKaRa bamhi digested puc19 plasmid
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Bamhi Digested Puc19 Plasmid, supplied by TaKaRa, used in various techniques. Bioz Stars score: 93/100, based on 15 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    TaKaRa puc19 t cloning kit
    Topological inactivation activities. Lane 1 indicated <t>pUC18</t> as a control; Lane 2 indicated the products of pUC18 <t>DNA</t> treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.
    Puc19 T Cloning Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    TaKaRa supercoiled puc19 plasmid dna
    Ability of AccGSTS1 to protect against <t>DNA</t> damage in the MFO system. Lane 0 <t>pUC19</t> plasmid DNA only; lane 1 pUC19 plasmid DNA + FeCl 3 ; lane 2 pUC19 plasmid DNA + FeCl 3 + dithiothreitol (DTT); lane 3 pUC19
    Supercoiled Puc19 Plasmid Dna, supplied by TaKaRa, used in various techniques. Bioz Stars score: 76/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    TaKaRa pcr linearized puc19 brca1ex18 vector
    Ability of AccGSTS1 to protect against <t>DNA</t> damage in the MFO system. Lane 0 <t>pUC19</t> plasmid DNA only; lane 1 pUC19 plasmid DNA + FeCl 3 ; lane 2 pUC19 plasmid DNA + FeCl 3 + dithiothreitol (DTT); lane 3 pUC19
    Pcr Linearized Puc19 Brca1ex18 Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 79/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    TaKaRa in fusion hd cloning kit
    Ability of AccGSTS1 to protect against <t>DNA</t> damage in the MFO system. Lane 0 <t>pUC19</t> plasmid DNA only; lane 1 pUC19 plasmid DNA + FeCl 3 ; lane 2 pUC19 plasmid DNA + FeCl 3 + dithiothreitol (DTT); lane 3 pUC19
    In Fusion Hd Cloning Kit, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 12308 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    QdNO-induced DNA strand breakage. (A, B) E . coli DH5α cells harboring the pBR322 plasmid were incubated with the indicated concentration of CYA (A) or OLA (B) at 37°C for 2 h. TPZ was used as the positive control drug. (C) Inhibition of the degradation of pUC-19 plasmid in E . coli DH5α by free radical scavengers. Bacteria harboring pUC-19 plasmid were incubated with 100 mM mannitol, thiourea, acetone, β-ME, ethanol, methanol and NaN 3 (from lane 2 to lane 8), and after 4 μg/ml CYA was added, the cells were incubated at 37°C for 2 h. The untreated bacteria (lane 1) and the bacteria treated only with CYA but no scavengers (lane 9) were used as the blank and the control, respectively. (D, E) Supercoiled pBR322 DNA (10 μg/ml) was incubated with the indicated concentration of CYA (D) or OLA (E) in the presence of xanthine oxidase/xanthine at 37°C for 0.5 h. The plasmids isolated from the treated bacteria (A-C) or the treated plasmids (D, E) were electrophoretically separated. XO/X, xanthine oxidase/xanthine; LM, linear DNA marker; SM, supercoiled DNA marker. SC, L and OC indicate supercoiled, linear and open circular DNA, respectively.

    Journal: PLoS ONE

    Article Title: Systematic and Molecular Basis of the Antibacterial Action of Quinoxaline 1,4-Di-N-Oxides against Escherichia coli

    doi: 10.1371/journal.pone.0136450

    Figure Lengend Snippet: QdNO-induced DNA strand breakage. (A, B) E . coli DH5α cells harboring the pBR322 plasmid were incubated with the indicated concentration of CYA (A) or OLA (B) at 37°C for 2 h. TPZ was used as the positive control drug. (C) Inhibition of the degradation of pUC-19 plasmid in E . coli DH5α by free radical scavengers. Bacteria harboring pUC-19 plasmid were incubated with 100 mM mannitol, thiourea, acetone, β-ME, ethanol, methanol and NaN 3 (from lane 2 to lane 8), and after 4 μg/ml CYA was added, the cells were incubated at 37°C for 2 h. The untreated bacteria (lane 1) and the bacteria treated only with CYA but no scavengers (lane 9) were used as the blank and the control, respectively. (D, E) Supercoiled pBR322 DNA (10 μg/ml) was incubated with the indicated concentration of CYA (D) or OLA (E) in the presence of xanthine oxidase/xanthine at 37°C for 0.5 h. The plasmids isolated from the treated bacteria (A-C) or the treated plasmids (D, E) were electrophoretically separated. XO/X, xanthine oxidase/xanthine; LM, linear DNA marker; SM, supercoiled DNA marker. SC, L and OC indicate supercoiled, linear and open circular DNA, respectively.

    Article Snippet: Supercoiled plasmid pBR322 and pUC-19 were purchased from Takara (Dalian, China).

    Techniques: Plasmid Preparation, Incubation, Concentration Assay, Positive Control, Inhibition, Isolation, Marker

    ttMutS2 preferably digested the DNA containing branched DNA structures. A , pUC19 or pUC(AT) was incubated with or without 5 n m DNase I. B , the same experiment as in A was performed with 0, 100, and 200 n m ttMutS2. The reaction solutions were incubated

    Journal:

    Article Title: Crystal Structure of MutS2 Endonuclease Domain and the Mechanism of Homologous Recombination Suppression *Crystal Structure of MutS2 Endonuclease Domain and the Mechanism of Homologous Recombination Suppression * S⃞

    doi: 10.1074/jbc.M806755200

    Figure Lengend Snippet: ttMutS2 preferably digested the DNA containing branched DNA structures. A , pUC19 or pUC(AT) was incubated with or without 5 n m DNase I. B , the same experiment as in A was performed with 0, 100, and 200 n m ttMutS2. The reaction solutions were incubated

    Article Snippet: As shown in , ttMutS2 digested pUC(AT) more preferably than pUC19, a normal plasmid DNA, whereas a negative control DNase I equally digested the two kinds of plasmid DNAs ( ).

    Techniques: Incubation

    The SMR domain of SOT1 has nuclease activity. ( A ) Purified MBP, SOT1 SMR , and Gm-SOT1 SMR used for nuclease activity were resolved by SDS/PAGE with Coomassie Brilliant Blue (CBB) staining or with the analyses of immunoblot using the MBP antibody. The marker sizes are shown to the Left . ( B ) DNA endonuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . MBP, SOT1 SMR , and Gm-SOT1 SMR at different concentrations were incubated with 5 ng pUC19 plasmid DNA at 25 °C for 60 min in the presence of 3 mM MgCl 2 . The reactions were stopped by loading buffer and were electrophoresed on 1.2% (wt/vol) agarose gels. Parallel experiments were carried out with EcoRI to linearize pUC19 or with H 2 O as a control. OC, open circular; CCC, covalently closed-circular forms of pUC19. ( C ) Effects of Mg 2+ , Ca 2+ , and Mn 2+ on DNA nicking activities of the SMR domains of SOT1 and Gm-SOT1 SMR . A total of 5 ng pUC19 was incubated with H 2 O, MgCl 2 , CaCl 2 , or MnCl 2 and 100 nM MBP, SOT1 SMR , and Gm-SOT1 SMR . The concentration of each cation was 3 mM. ( D ) Analyses of DNA nicking activity of Gm-SOT1. MBP and Gm-SOT1 were incubated with 5 ng pUC19 plasmid DNA at 25 °C for 60 min in the presence of 3 mM MgCl 2 . ( E ) RNA nuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . MBP, SOT1 SMR , and Gm-SOT1 SMR with different concentrations were incubated with total wild-type Arabidopsis RNA at 25 °C for 30 min. The reaction products were separated in agarose/formaldehyde gels and observed by ethidium bromide staining. ( F ) Effects of Mg 2+ , Ca 2+ , and Mn 2+ on RNA nuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . A total of 100 nM MBP, SOT1 SMR , and Gm-SOT1 SMR were incubated with total wild-type Arabidopsis RNA in the presence of MgCl 2 , CaCl 2 , or MnCl 2 . The concentration of each cation was 3 mM. ( G ) Accumulation of total RNAs in WT, sot1-3/35S:SOT1 SMR -HA , and sot1-3 plants. A total of 3 μg total RNAs from 12-d-old WT, sot1-3/35S:SOT1 SMR -HA , and sot1-3 seedlings were separated in agarose/formaldehyde gels and observed by ethidium bromide staining. ( H ) Analyses of RNA nuclease activity of Gm-SOT1. MBP and Gm-SOT1 with different concentrations were incubated with total wild-type Arabidopsis RNA at 25 °C for 30 min. The reaction products were detected by electrophoretic separation in agarose/formaldehyde gels with ethidium bromide staining.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: PPR-SMR protein SOT1 has RNA endonuclease activity

    doi: 10.1073/pnas.1612460114

    Figure Lengend Snippet: The SMR domain of SOT1 has nuclease activity. ( A ) Purified MBP, SOT1 SMR , and Gm-SOT1 SMR used for nuclease activity were resolved by SDS/PAGE with Coomassie Brilliant Blue (CBB) staining or with the analyses of immunoblot using the MBP antibody. The marker sizes are shown to the Left . ( B ) DNA endonuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . MBP, SOT1 SMR , and Gm-SOT1 SMR at different concentrations were incubated with 5 ng pUC19 plasmid DNA at 25 °C for 60 min in the presence of 3 mM MgCl 2 . The reactions were stopped by loading buffer and were electrophoresed on 1.2% (wt/vol) agarose gels. Parallel experiments were carried out with EcoRI to linearize pUC19 or with H 2 O as a control. OC, open circular; CCC, covalently closed-circular forms of pUC19. ( C ) Effects of Mg 2+ , Ca 2+ , and Mn 2+ on DNA nicking activities of the SMR domains of SOT1 and Gm-SOT1 SMR . A total of 5 ng pUC19 was incubated with H 2 O, MgCl 2 , CaCl 2 , or MnCl 2 and 100 nM MBP, SOT1 SMR , and Gm-SOT1 SMR . The concentration of each cation was 3 mM. ( D ) Analyses of DNA nicking activity of Gm-SOT1. MBP and Gm-SOT1 were incubated with 5 ng pUC19 plasmid DNA at 25 °C for 60 min in the presence of 3 mM MgCl 2 . ( E ) RNA nuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . MBP, SOT1 SMR , and Gm-SOT1 SMR with different concentrations were incubated with total wild-type Arabidopsis RNA at 25 °C for 30 min. The reaction products were separated in agarose/formaldehyde gels and observed by ethidium bromide staining. ( F ) Effects of Mg 2+ , Ca 2+ , and Mn 2+ on RNA nuclease activities of the SMR domains of SOT1 and Gm-SOT1 SMR . A total of 100 nM MBP, SOT1 SMR , and Gm-SOT1 SMR were incubated with total wild-type Arabidopsis RNA in the presence of MgCl 2 , CaCl 2 , or MnCl 2 . The concentration of each cation was 3 mM. ( G ) Accumulation of total RNAs in WT, sot1-3/35S:SOT1 SMR -HA , and sot1-3 plants. A total of 3 μg total RNAs from 12-d-old WT, sot1-3/35S:SOT1 SMR -HA , and sot1-3 seedlings were separated in agarose/formaldehyde gels and observed by ethidium bromide staining. ( H ) Analyses of RNA nuclease activity of Gm-SOT1. MBP and Gm-SOT1 with different concentrations were incubated with total wild-type Arabidopsis RNA at 25 °C for 30 min. The reaction products were detected by electrophoretic separation in agarose/formaldehyde gels with ethidium bromide staining.

    Article Snippet: The pUC19 (Takara) plasmid DNA (5 ng μL−1 ) was incubated with MBP, SOT1SMR , SOT1SMR variants, Gm-SOT1SMR , Gm-SOT1SMR variants, or Gm-SOT1 in buffer [20 mM phosphate, pH 7.5, 160 mM NaCl, 40 mM KCl, and 4% (vol/vol) glycerol] at 25 °C for 60 min.

    Techniques: Activity Assay, Purification, SDS Page, Staining, Marker, Incubation, Plasmid Preparation, Countercurrent Chromatography, Concentration Assay

    Comparison of expression levels of the AsnO protein between lac and T7 promoters by SDS-PAGE analysis. Lane 1, molecular mass standards; lanes 2 and 3, lac expression system using the pUC19 vector; lanes 4 and 5, T7 expression system using the pET-21a(+)

    Journal: Applied and Environmental Microbiology

    Article Title: One-Pot Production of l-threo-3-Hydroxyaspartic Acid Using Asparaginase-Deficient Escherichia coli Expressing Asparagine Hydroxylase of Streptomyces coelicolor A3(2)

    doi: 10.1128/AEM.03963-14

    Figure Lengend Snippet: Comparison of expression levels of the AsnO protein between lac and T7 promoters by SDS-PAGE analysis. Lane 1, molecular mass standards; lanes 2 and 3, lac expression system using the pUC19 vector; lanes 4 and 5, T7 expression system using the pET-21a(+)

    Article Snippet: E. coli strains JW1756 and JW2924 were obtained from the National Bio Resource Project (NBRP) (National Institute of Genetics, Japan) ( ) and used for whole-cell reactions together with pUC19-based vectors (TaKaRa Bio, Shiga, Japan).

    Techniques: Expressing, SDS Page, Plasmid Preparation, Positron Emission Tomography

    Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from pUC19 were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Tetrahymena proteins p80 and p95 are not core telomerase components

    doi: 10.1073/pnas.221456398

    Figure Lengend Snippet: Interactions of p80 with various RNAs. p80 and tTERT were expressed individually by in vitro transcription and translation in the presence of [ 35 S]methionine using rabbit reticulocyte lysates. DNA templates for T7 transcription of telomerase RNA, 7SL RNA, U2 RNA, and a random RNA derived from pUC19 were included in the reaction. Immunoprecipitations were performed with the p80-R and TERT-P antibodies. ( A ) Northern analysis to detect telomerase RNA, 7SL RNA, and random RNA from immunoprecipitations. ( B ) Northern analysis to detect telomerase RNA and U2 RNA from immunoprecipitations. ( C ) Autoradiography of immunoprecipitated 35 S-methionine-labeled proteins after SDS/PAGE.

    Article Snippet: A random RNA sequence for T7 transcription was generated from a PCR template derived from the plasmid pUC19 (CLONTECH).

    Techniques: In Vitro, Derivative Assay, Northern Blot, Autoradiography, Immunoprecipitation, Labeling, SDS Page

    Construction strategy of pIC-2DuCV. a Two full-length genomes of DuCV strain GH01, denoted IC1 and IC2, were amplified. b IC1 and IC2 were ligated into the pUC19 vector to yield pIC-1 and pIC-2, respectively. c IC-2 was ligated head-to-tail to pIC-1 to produce a tandem-dimerized DuCV DNA clone, which was denoted pIC-2DuCV. d pIC-2DuCV

    Journal: Virology Journal

    Article Title: Rescue of a duck circovirus from an infectious DNA clone in ducklings

    doi: 10.1186/s12985-015-0312-6

    Figure Lengend Snippet: Construction strategy of pIC-2DuCV. a Two full-length genomes of DuCV strain GH01, denoted IC1 and IC2, were amplified. b IC1 and IC2 were ligated into the pUC19 vector to yield pIC-1 and pIC-2, respectively. c IC-2 was ligated head-to-tail to pIC-1 to produce a tandem-dimerized DuCV DNA clone, which was denoted pIC-2DuCV. d pIC-2DuCV

    Article Snippet: The recombinant pIC-2DuCV and pIC-Mu2DuCV plasmids and the pUC19 vector were inoculated intramuscularly into five ducklings.

    Techniques: Amplification, Plasmid Preparation

    Construction strategy of pIC-Mu2DuCV. a Two full-length genomes of DuCV strain GH01, denoted IC-Mu1 and IC-Mu2, were amplified by overlapping PCR. b IC-Mu1 and IC-Mu2 were ligated into the pUC19 vector to yield pIC-Mu1 and pIC-Mu2, respectively. c IC-Mu2 was ligated head-to-tail to pIC-Mu1 to produce a tandem-dimerized DuCV DNA clone, which was denoted pIC-Mu2DuCV. d pIC-Mu2DuCV

    Journal: Virology Journal

    Article Title: Rescue of a duck circovirus from an infectious DNA clone in ducklings

    doi: 10.1186/s12985-015-0312-6

    Figure Lengend Snippet: Construction strategy of pIC-Mu2DuCV. a Two full-length genomes of DuCV strain GH01, denoted IC-Mu1 and IC-Mu2, were amplified by overlapping PCR. b IC-Mu1 and IC-Mu2 were ligated into the pUC19 vector to yield pIC-Mu1 and pIC-Mu2, respectively. c IC-Mu2 was ligated head-to-tail to pIC-Mu1 to produce a tandem-dimerized DuCV DNA clone, which was denoted pIC-Mu2DuCV. d pIC-Mu2DuCV

    Article Snippet: The recombinant pIC-2DuCV and pIC-Mu2DuCV plasmids and the pUC19 vector were inoculated intramuscularly into five ducklings.

    Techniques: Amplification, Polymerase Chain Reaction, Plasmid Preparation

    Identification of recombinant pIC-Mu2DuCV plasmid by restriction enzyme digestion. M, wide-range DNA marker (500 ~ 12,000); 1, digested with Hind III; 2, digested with Hind III and BamH I; 3, digested with BamH I and EcoR I; 4, digested with Hind III and EcoR I; 5, digested with Xho I; 6, pUC19 digested with Hind III; 7, pUC19

    Journal: Virology Journal

    Article Title: Rescue of a duck circovirus from an infectious DNA clone in ducklings

    doi: 10.1186/s12985-015-0312-6

    Figure Lengend Snippet: Identification of recombinant pIC-Mu2DuCV plasmid by restriction enzyme digestion. M, wide-range DNA marker (500 ~ 12,000); 1, digested with Hind III; 2, digested with Hind III and BamH I; 3, digested with BamH I and EcoR I; 4, digested with Hind III and EcoR I; 5, digested with Xho I; 6, pUC19 digested with Hind III; 7, pUC19

    Article Snippet: The recombinant pIC-2DuCV and pIC-Mu2DuCV plasmids and the pUC19 vector were inoculated intramuscularly into five ducklings.

    Techniques: Recombinant, Plasmid Preparation, Marker

    Identification of the recombinant plasmid pIC-2DuCV by restriction enzyme digestion. M, wide-range DNA marker (500 ~ 12,000); 1, digested with Hind III; 2, digested with Hind III and BamH I; 3, digested with BamH I and EcoR I; 4, digested with Hind III and EcoR I; 5, pUC19 digested with Hind III

    Journal: Virology Journal

    Article Title: Rescue of a duck circovirus from an infectious DNA clone in ducklings

    doi: 10.1186/s12985-015-0312-6

    Figure Lengend Snippet: Identification of the recombinant plasmid pIC-2DuCV by restriction enzyme digestion. M, wide-range DNA marker (500 ~ 12,000); 1, digested with Hind III; 2, digested with Hind III and BamH I; 3, digested with BamH I and EcoR I; 4, digested with Hind III and EcoR I; 5, pUC19 digested with Hind III

    Article Snippet: The recombinant pIC-2DuCV and pIC-Mu2DuCV plasmids and the pUC19 vector were inoculated intramuscularly into five ducklings.

    Techniques: Recombinant, Plasmid Preparation, Marker

    Validation of ABI-REC through double-resistance reporter assay. ( A ) The design of double resistance reporter assay. The fusion of the 1.6 kb kan R cassette (from pIRES2-EGFP) with the 2.7 kb pUC19 plasmid (ampicillin resistance) renders the new recombinant plasmid resistant to both kanamycin and ampicillin. The plasmid grows on (Kan + Amp) LB plates. An artificial ApaI restriction site is introduced into the 5′ fusion site to precisely localize the insertion site. ( B ) An asymmetric bridge PCR efficiently fuses the Kan R cassette into the pUC19 plasmid and generates a hybrid fragment. The gradient concentration of the primer pR was assessed and identified as within the optimal range from 2 nM to 0.2 nM in the bridge PCR. A conventional two-primer PCR reaction was conducted in parallel to compare their amplification efficiency. This indicates that the quantity of pR primer is critical to the output of fused sequence in the bridge PCR reaction. ( C ) Double resistant colonies were found to grow on (Kan + Amp) LB plates. Bridge PCR products, purified fused sequences, two-primer PCR products (extension), and a mixture of purified Kan R cassettes with pUC19 (no extension) were transformed into DH5α competent cells. Double resistant colonies were present for bridge PCR products and purified fused sequence, whereas the latter two did not produce viable colonies. This implies that intramolecular recombination occurs within the fused sequence, producing double resistant plasmids that renders cells resistant to both kanamycin and ampicillin. pUC19 and pIRES2-EGFP plasmids were not able to grow on the double-drug plates, precluding the risk of random integration of Kan R cassette into the E.coli genome. ( D ) Plasmids of the single colonies in (Kan + Amp) LB plates were extracted and digested using SalI and ApaI. As shown in the electrophoresis gel, the 1.6 kb insert was released, indicating that the Kan R cassette had been fused into pUC19 at pre-determined site. ( E ) Sequencing of the single colonies revealed the insertion of the Kan R cassette and the presence of artificial ApaI restriction site, further proving that the insert of choice has been fused with target plasmid in a restriction- and ligase-free manner. The red rectangle represents the restriction sites.

    Journal: BMC Biotechnology

    Article Title: An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy

    doi: 10.1186/1472-6750-12-39

    Figure Lengend Snippet: Validation of ABI-REC through double-resistance reporter assay. ( A ) The design of double resistance reporter assay. The fusion of the 1.6 kb kan R cassette (from pIRES2-EGFP) with the 2.7 kb pUC19 plasmid (ampicillin resistance) renders the new recombinant plasmid resistant to both kanamycin and ampicillin. The plasmid grows on (Kan + Amp) LB plates. An artificial ApaI restriction site is introduced into the 5′ fusion site to precisely localize the insertion site. ( B ) An asymmetric bridge PCR efficiently fuses the Kan R cassette into the pUC19 plasmid and generates a hybrid fragment. The gradient concentration of the primer pR was assessed and identified as within the optimal range from 2 nM to 0.2 nM in the bridge PCR. A conventional two-primer PCR reaction was conducted in parallel to compare their amplification efficiency. This indicates that the quantity of pR primer is critical to the output of fused sequence in the bridge PCR reaction. ( C ) Double resistant colonies were found to grow on (Kan + Amp) LB plates. Bridge PCR products, purified fused sequences, two-primer PCR products (extension), and a mixture of purified Kan R cassettes with pUC19 (no extension) were transformed into DH5α competent cells. Double resistant colonies were present for bridge PCR products and purified fused sequence, whereas the latter two did not produce viable colonies. This implies that intramolecular recombination occurs within the fused sequence, producing double resistant plasmids that renders cells resistant to both kanamycin and ampicillin. pUC19 and pIRES2-EGFP plasmids were not able to grow on the double-drug plates, precluding the risk of random integration of Kan R cassette into the E.coli genome. ( D ) Plasmids of the single colonies in (Kan + Amp) LB plates were extracted and digested using SalI and ApaI. As shown in the electrophoresis gel, the 1.6 kb insert was released, indicating that the Kan R cassette had been fused into pUC19 at pre-determined site. ( E ) Sequencing of the single colonies revealed the insertion of the Kan R cassette and the presence of artificial ApaI restriction site, further proving that the insert of choice has been fused with target plasmid in a restriction- and ligase-free manner. The red rectangle represents the restriction sites.

    Article Snippet: The pUC19 cloning plasmid was from Takara (Japan). pIRES2-EGFP was purchased from Clontech (U.S.).

    Techniques: Reporter Assay, Plasmid Preparation, Recombinant, Bridge PCR, Concentration Assay, Polymerase Chain Reaction, Amplification, Sequencing, Purification, Transformation Assay, Electrophoresis

    Effects of homology and insert length upon the efficiency of ABI-REC. ( A ) Homology was varied by increasing the length of P1R primer in the asymmetric bridge PCR reaction. The reactions with 20 bp and 25 bp P1R primers gave rise to maximum output of fused fragments. ( B ) Transformation of equal volumes of these PCR products into E.coli cells produced double resistant colonies with various numbers. Colony capacity varied, with colonies producing maximum target fragment output being more numerous. The number of colonies per plate was plotted against homology. Error bars indicate mean ± SD from three independent assays. ( C ) Inserts ranging from 1.6 kb to 4 kb were fused into pUC19 in asymmetric PCR reactions. Red arrows denote the fused fragments. ( D ) Transformation of equal volumes of these PCR products into E.coli cells produced various numbers of double resistant colonies. Quantitation of single colonies revealed that colony capacity varied, with colonies producing maximum target fragment output being more numerous. The number of colonies per plate was plotted against insert size. Error bars indicate mean ± SD from three independent assays. ( E ) ApaI and SalI restriction digestion of the plasmids extracted from the single colonies in ( D ). All plasmids released the inserts as designed. Please note that 2.5 kb insert is nearly identical to backbone pUC19, as one band was observed. In addition, the 4 kb insert has three ApaI sites, one of which is only 133 bp in size and therefore undetectable in the gel. A 367 bp band is denoted by a red arrow.

    Journal: BMC Biotechnology

    Article Title: An asymmetric PCR-based, reliable and rapid single-tube native DNA engineering strategy

    doi: 10.1186/1472-6750-12-39

    Figure Lengend Snippet: Effects of homology and insert length upon the efficiency of ABI-REC. ( A ) Homology was varied by increasing the length of P1R primer in the asymmetric bridge PCR reaction. The reactions with 20 bp and 25 bp P1R primers gave rise to maximum output of fused fragments. ( B ) Transformation of equal volumes of these PCR products into E.coli cells produced double resistant colonies with various numbers. Colony capacity varied, with colonies producing maximum target fragment output being more numerous. The number of colonies per plate was plotted against homology. Error bars indicate mean ± SD from three independent assays. ( C ) Inserts ranging from 1.6 kb to 4 kb were fused into pUC19 in asymmetric PCR reactions. Red arrows denote the fused fragments. ( D ) Transformation of equal volumes of these PCR products into E.coli cells produced various numbers of double resistant colonies. Quantitation of single colonies revealed that colony capacity varied, with colonies producing maximum target fragment output being more numerous. The number of colonies per plate was plotted against insert size. Error bars indicate mean ± SD from three independent assays. ( E ) ApaI and SalI restriction digestion of the plasmids extracted from the single colonies in ( D ). All plasmids released the inserts as designed. Please note that 2.5 kb insert is nearly identical to backbone pUC19, as one band was observed. In addition, the 4 kb insert has three ApaI sites, one of which is only 133 bp in size and therefore undetectable in the gel. A 367 bp band is denoted by a red arrow.

    Article Snippet: The pUC19 cloning plasmid was from Takara (Japan). pIRES2-EGFP was purchased from Clontech (U.S.).

    Techniques: Bridge PCR, Transformation Assay, Polymerase Chain Reaction, Produced, Quantitation Assay

    Topological inactivation activities. Lane 1 indicated pUC18 as a control; Lane 2 indicated the products of pUC18 DNA treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.

    Journal: PLoS ONE

    Article Title: A Novel Method for Simultaneous Production of Two Ribosome-Inactivating Proteins, ?-MMC and MAP30, from Momordica charantia L

    doi: 10.1371/journal.pone.0101998

    Figure Lengend Snippet: Topological inactivation activities. Lane 1 indicated pUC18 as a control; Lane 2 indicated the products of pUC18 DNA treated with α-MMC; Lane M indicated DNA ladder; Lane 3 indicated the products of pUC18 DNA treated with MAP30.

    Article Snippet: LMW Calibration Kit was supplied by SIBAS (Shanghai, China). pUC18 DNA used in detection of topological activity was obtained from TAKARA (Dalian, China).

    Techniques:

    Ability of AccGSTS1 to protect against DNA damage in the MFO system. Lane 0 pUC19 plasmid DNA only; lane 1 pUC19 plasmid DNA + FeCl 3 ; lane 2 pUC19 plasmid DNA + FeCl 3 + dithiothreitol (DTT); lane 3 pUC19

    Journal: Cell Stress & Chaperones

    Article Title: Identification, genomic organization, and oxidative stress response of a sigma class glutathione S-transferase gene (AccGSTS1) in the honey bee, Apis cerana cerana

    doi: 10.1007/s12192-012-0394-7

    Figure Lengend Snippet: Ability of AccGSTS1 to protect against DNA damage in the MFO system. Lane 0 pUC19 plasmid DNA only; lane 1 pUC19 plasmid DNA + FeCl 3 ; lane 2 pUC19 plasmid DNA + FeCl 3 + dithiothreitol (DTT); lane 3 pUC19

    Article Snippet: The reaction mixture (25 μl) containing 2 μg of supercoiled pUC19 plasmid DNA (TaKaRa, Dalian, China), 3 μM FeCl3 and 10 mM DTT in 100 mM Hepes buffer (pH 7.0) and increasing concentrations of AccGSTS1 was incubated at 37 °C for 3 h and was then subjected to 1 % agarose gel electrophoresis for the determination of DNA cleavage.

    Techniques: Plasmid Preparation