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p53 dbss  (TaKaRa)


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    TaKaRa p53 dbss
    The effect of suppressor amino acid changes on <t>p53</t> cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the <t>p53</t> <t>expression</t> plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.
    P53 Dbss, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p53 dbss/product/TaKaRa
    Average 86 stars, based on 1 article reviews
    p53 dbss - by Bioz Stars, 2026-02
    86/100 stars

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    1) Product Images from "A global suppressor motif for p53 cancer mutants"

    Article Title: A global suppressor motif for p53 cancer mutants

    Journal:

    doi: 10.1073/pnas.0401162101

    The effect of suppressor amino acid changes on p53 cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the p53 expression plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.
    Figure Legend Snippet: The effect of suppressor amino acid changes on p53 cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the p53 expression plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.

    Techniques Used: Mutagenesis, Activity Assay, Expressing, Incubation, Clone Assay, Selection

    The global 235-239-240 suppressor motif rescues many common p53 cancer mutants in mammalian reporter gene assays for single p53 DBSs. The transcriptional activity of p53 cancer mutants with and without suppressor amino acids was evaluated in transient reporter gene assays in p53-negative H1299 cells. Luciferase activity, adjusted for transfection efficiency by using Renilla luciferase activity, was determined 24 h after transfection. The adjusted luciferase activity of cell lysates transfected with reporter plasmids and WT p53 expression plasmid was set as 100%. Shown are the mean and SD for three independent experiments. The reporter gene plasmids contained the DBSs of the p53 target genes GML (gray bars) or KILLER/DR5 (black bars) in the context of a heterologous yeast promoter. p53 cancer mutants are written out, and p53 mutants with cancer and suppressor amino acids are indicated by numbers that can be crossreferenced with Table 2 and Fig. 1C. The majority of p53 cancer mutants was rescued by at least one suppressor combination with transcriptional activities ranging from 40% to 130% of WT p53 activity. In the case of R158L, V173M, Y205C, and Y220C, the presence of suppressor amino acids resulted in only 20% of WT p53 activity. However, this result still represents a significant rescue effect, considering the lack of activity of the p53 cancer mutants alone.
    Figure Legend Snippet: The global 235-239-240 suppressor motif rescues many common p53 cancer mutants in mammalian reporter gene assays for single p53 DBSs. The transcriptional activity of p53 cancer mutants with and without suppressor amino acids was evaluated in transient reporter gene assays in p53-negative H1299 cells. Luciferase activity, adjusted for transfection efficiency by using Renilla luciferase activity, was determined 24 h after transfection. The adjusted luciferase activity of cell lysates transfected with reporter plasmids and WT p53 expression plasmid was set as 100%. Shown are the mean and SD for three independent experiments. The reporter gene plasmids contained the DBSs of the p53 target genes GML (gray bars) or KILLER/DR5 (black bars) in the context of a heterologous yeast promoter. p53 cancer mutants are written out, and p53 mutants with cancer and suppressor amino acids are indicated by numbers that can be crossreferenced with Table 2 and Fig. 1C. The majority of p53 cancer mutants was rescued by at least one suppressor combination with transcriptional activities ranging from 40% to 130% of WT p53 activity. In the case of R158L, V173M, Y205C, and Y220C, the presence of suppressor amino acids resulted in only 20% of WT p53 activity. However, this result still represents a significant rescue effect, considering the lack of activity of the p53 cancer mutants alone.

    Techniques Used: Activity Assay, Luciferase, Transfection, Expressing, Plasmid Preparation



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    p53 dbss  (TaKaRa)
    86
    TaKaRa p53 dbss
    The effect of suppressor amino acid changes on <t>p53</t> cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the <t>p53</t> <t>expression</t> plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.
    P53 Dbss, supplied by TaKaRa, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/p53 dbss/product/TaKaRa
    Average 86 stars, based on 1 article reviews
    p53 dbss - by Bioz Stars, 2026-02
    86/100 stars
    		  Buy from Supplier

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    The effect of suppressor amino acid changes on p53 cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the p53 expression plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.

    Journal:

    Article Title: A global suppressor motif for p53 cancer mutants

    doi: 10.1073/pnas.0401162101

    Figure Lengend Snippet: The effect of suppressor amino acid changes on p53 cancer mutants in the p53 yeast assay. (A) PCR-mediated mutagenesis yielded suppressor mutation combinations that rescued transcriptional activity of four of the eight most common p53 cancer mutants, G245S, R249S, R273C, and R273H. SC-His plates select for the p53 expression plasmids and SC-Ura plates select for transcriptionally active p53. Pictures were taken after 2 days of incubation at 30°C. All yeast clones are represented by two patches. p53 cancer mutants are named (e.g., G245S), p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 1. (B) Design of the oligonucleotide-based mutagenesis. Expression plasmids for 30 of the most common p53 cancer mutants were gapped by using PflMI and NsiI. Pairs of annealed oligonucleotides representing all amino acid changes at codons 239 or 240 were cloned into the gap. The design also included background mutagenesis of the remaining codons between 225 and 241 that resulted in approximately one misincorporation per 100 nucleotides. (C) The global suppressor motif 235-239-240 rescues 16 of 30 of the most common p53 cancer mutants in the yeast p53 assay. All yeast transformants were processed and are shown as described for A. V272M was Ura+, but less so than V272M plus N239W. Identification of V272M plus N239W was likely possible, because the selection for rescued p53 cancer mutants (plating of transformants directly onto SC-His-Ura plates) was more stringent than subsequent confirmation of phenotypes for yeast patches by replica plating. V272M was transcriptionally inactive in mammalian reporter gene assays, whereas V272M plus N239W showed transcriptional activity (see Figs. ​Figs.22 and ​and3).3). p53 mutants with cancer and suppressor combinations are numbered, and are crossreferenced with Table 2.

    Article Snippet: Reporter gene plasmids with single p53 DBSs were based on pp53-TA-Luc (Clontech; ref 20 ).

    Techniques: Mutagenesis, Activity Assay, Expressing, Incubation, Clone Assay, Selection

    The global 235-239-240 suppressor motif rescues many common p53 cancer mutants in mammalian reporter gene assays for single p53 DBSs. The transcriptional activity of p53 cancer mutants with and without suppressor amino acids was evaluated in transient reporter gene assays in p53-negative H1299 cells. Luciferase activity, adjusted for transfection efficiency by using Renilla luciferase activity, was determined 24 h after transfection. The adjusted luciferase activity of cell lysates transfected with reporter plasmids and WT p53 expression plasmid was set as 100%. Shown are the mean and SD for three independent experiments. The reporter gene plasmids contained the DBSs of the p53 target genes GML (gray bars) or KILLER/DR5 (black bars) in the context of a heterologous yeast promoter. p53 cancer mutants are written out, and p53 mutants with cancer and suppressor amino acids are indicated by numbers that can be crossreferenced with Table 2 and Fig. 1C. The majority of p53 cancer mutants was rescued by at least one suppressor combination with transcriptional activities ranging from 40% to 130% of WT p53 activity. In the case of R158L, V173M, Y205C, and Y220C, the presence of suppressor amino acids resulted in only 20% of WT p53 activity. However, this result still represents a significant rescue effect, considering the lack of activity of the p53 cancer mutants alone.

    Journal:

    Article Title: A global suppressor motif for p53 cancer mutants

    doi: 10.1073/pnas.0401162101

    Figure Lengend Snippet: The global 235-239-240 suppressor motif rescues many common p53 cancer mutants in mammalian reporter gene assays for single p53 DBSs. The transcriptional activity of p53 cancer mutants with and without suppressor amino acids was evaluated in transient reporter gene assays in p53-negative H1299 cells. Luciferase activity, adjusted for transfection efficiency by using Renilla luciferase activity, was determined 24 h after transfection. The adjusted luciferase activity of cell lysates transfected with reporter plasmids and WT p53 expression plasmid was set as 100%. Shown are the mean and SD for three independent experiments. The reporter gene plasmids contained the DBSs of the p53 target genes GML (gray bars) or KILLER/DR5 (black bars) in the context of a heterologous yeast promoter. p53 cancer mutants are written out, and p53 mutants with cancer and suppressor amino acids are indicated by numbers that can be crossreferenced with Table 2 and Fig. 1C. The majority of p53 cancer mutants was rescued by at least one suppressor combination with transcriptional activities ranging from 40% to 130% of WT p53 activity. In the case of R158L, V173M, Y205C, and Y220C, the presence of suppressor amino acids resulted in only 20% of WT p53 activity. However, this result still represents a significant rescue effect, considering the lack of activity of the p53 cancer mutants alone.

    Article Snippet: Reporter gene plasmids with single p53 DBSs were based on pp53-TA-Luc (Clontech; ref 20 ).

    Techniques: Activity Assay, Luciferase, Transfection, Expressing, Plasmid Preparation