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pas2 1 bait vector  (TaKaRa)


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    TaKaRa pas2 1 bait vector
    Pas2 1 Bait Vector, supplied by TaKaRa, used in various techniques. Bioz Stars score: 96/100, based on 619 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/bait+vector+pas2+1/pmCherry+Vector/pmc04633242-35-67-70
    Average 96 stars, based on 619 article reviews
    pas2 1 bait vector - by Bioz Stars, 2026-09
    96/100 stars

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    Related Articles

    Polymerase Chain Reaction:

    Article Title: Nm23-H2 Interacts with a G Protein-coupled Receptor to Regulate Its Endocytosis through an Rac1-dependent Mechanism
    Article Snippet: Plasmids Construction—The TP C-terminal (TP CT) coding sequences (amino acids 328–407) were isolated by PCR using the Expand High Fidelity System (Roche Applied Science) with the following oligonucleotides TP CTF (5 -CAGGGATCCTCAATCCTTTCTGGACAGAGCCTTCCC-3 ) and TP CTR (5 -CAGGAATTCCGGAGTCTCACTCTGTGGCCCAGC-3 ). .. The PCR fragment was digested with EcoRI and XhoI and ligated into the bait vector pAS2-1 (Clontech) digested with the same enzymes and subsequently used to perform yeast two-hybrid screening as described below. pcDNA3-Myc-Nm23-H2WT, pcDNA3-Myc-Nm23-H2H118C, and pcDNA3-Nm23-H2- Kpn were prepared by PCR using the pGad424-Nm23-H2 plasmid (isolated by yeast two-hybrid screening) and the following primers, respectively, NM23F (5 -GAGGAATTCATGGCCAACCTGGAGCGCACC-3 ), NM23R (5 -GCGCTCGAGTCACAGATCCTCTTCAGAGATGAGTTTCTGTTCTTCATAGACCCAGTCATG-3 ), Nm23H118CF (5 -GGCAGGAACATCATTTGTGGCAGTGATTCAG-3 ), NM23H118CR (5 -CTGAATCACTGCCACAAAGATGTTCCTGCC-3 ), NM23 KpnF (5 -CTGAATCACTGCATGAATGATGGTCTCCCCAAGCATCACTCGGCC-3 ), and NM23 KpnR (5 -GGCCGAGTGATGCTTGGGGAGACCATCATTCATGGCAGTGATTCAG-3 ). ..

    Plasmid Preparation:

    Article Title: Nm23-H2 Interacts with a G Protein-coupled Receptor to Regulate Its Endocytosis through an Rac1-dependent Mechanism
    Article Snippet: Plasmids Construction—The TP C-terminal (TP CT) coding sequences (amino acids 328–407) were isolated by PCR using the Expand High Fidelity System (Roche Applied Science) with the following oligonucleotides TP CTF (5 -CAGGGATCCTCAATCCTTTCTGGACAGAGCCTTCCC-3 ) and TP CTR (5 -CAGGAATTCCGGAGTCTCACTCTGTGGCCCAGC-3 ). .. The PCR fragment was digested with EcoRI and XhoI and ligated into the bait vector pAS2-1 (Clontech) digested with the same enzymes and subsequently used to perform yeast two-hybrid screening as described below. pcDNA3-Myc-Nm23-H2WT, pcDNA3-Myc-Nm23-H2H118C, and pcDNA3-Nm23-H2- Kpn were prepared by PCR using the pGad424-Nm23-H2 plasmid (isolated by yeast two-hybrid screening) and the following primers, respectively, NM23F (5 -GAGGAATTCATGGCCAACCTGGAGCGCACC-3 ), NM23R (5 -GCGCTCGAGTCACAGATCCTCTTCAGAGATGAGTTTCTGTTCTTCATAGACCCAGTCATG-3 ), Nm23H118CF (5 -GGCAGGAACATCATTTGTGGCAGTGATTCAG-3 ), NM23H118CR (5 -CTGAATCACTGCCACAAAGATGTTCCTGCC-3 ), NM23 KpnF (5 -CTGAATCACTGCATGAATGATGGTCTCCCCAAGCATCACTCGGCC-3 ), and NM23 KpnR (5 -GGCCGAGTGATGCTTGGGGAGACCATCATTCATGGCAGTGATTCAG-3 ). ..

    Article Title: Virion-Wide Protein Interactions of Kaposi's Sarcoma-Associated Herpesvirus
    Article Snippet: .. The cDNAs for 11 tegument proteins or fragments were cloned into a Clontech Matchmaker system two-bait vector pAS2-1, which allows expression of a fusion protein of the DNA binding domain of yeast GAL4 protein and a tegument protein. cDNAs of all KSHV virion proteins (capsid, tegument, and envelope glycoproteins) were cloned into Matchmaker prey vector pACT2, generating the prey array listed in Table 1. ..

    Two Hybrid Screening:

    Article Title: Nm23-H2 Interacts with a G Protein-coupled Receptor to Regulate Its Endocytosis through an Rac1-dependent Mechanism
    Article Snippet: Plasmids Construction—The TP C-terminal (TP CT) coding sequences (amino acids 328–407) were isolated by PCR using the Expand High Fidelity System (Roche Applied Science) with the following oligonucleotides TP CTF (5 -CAGGGATCCTCAATCCTTTCTGGACAGAGCCTTCCC-3 ) and TP CTR (5 -CAGGAATTCCGGAGTCTCACTCTGTGGCCCAGC-3 ). .. The PCR fragment was digested with EcoRI and XhoI and ligated into the bait vector pAS2-1 (Clontech) digested with the same enzymes and subsequently used to perform yeast two-hybrid screening as described below. pcDNA3-Myc-Nm23-H2WT, pcDNA3-Myc-Nm23-H2H118C, and pcDNA3-Nm23-H2- Kpn were prepared by PCR using the pGad424-Nm23-H2 plasmid (isolated by yeast two-hybrid screening) and the following primers, respectively, NM23F (5 -GAGGAATTCATGGCCAACCTGGAGCGCACC-3 ), NM23R (5 -GCGCTCGAGTCACAGATCCTCTTCAGAGATGAGTTTCTGTTCTTCATAGACCCAGTCATG-3 ), Nm23H118CF (5 -GGCAGGAACATCATTTGTGGCAGTGATTCAG-3 ), NM23H118CR (5 -CTGAATCACTGCCACAAAGATGTTCCTGCC-3 ), NM23 KpnF (5 -CTGAATCACTGCATGAATGATGGTCTCCCCAAGCATCACTCGGCC-3 ), and NM23 KpnR (5 -GGCCGAGTGATGCTTGGGGAGACCATCATTCATGGCAGTGATTCAG-3 ). ..

    Isolation:

    Article Title: Nm23-H2 Interacts with a G Protein-coupled Receptor to Regulate Its Endocytosis through an Rac1-dependent Mechanism
    Article Snippet: Plasmids Construction—The TP C-terminal (TP CT) coding sequences (amino acids 328–407) were isolated by PCR using the Expand High Fidelity System (Roche Applied Science) with the following oligonucleotides TP CTF (5 -CAGGGATCCTCAATCCTTTCTGGACAGAGCCTTCCC-3 ) and TP CTR (5 -CAGGAATTCCGGAGTCTCACTCTGTGGCCCAGC-3 ). .. The PCR fragment was digested with EcoRI and XhoI and ligated into the bait vector pAS2-1 (Clontech) digested with the same enzymes and subsequently used to perform yeast two-hybrid screening as described below. pcDNA3-Myc-Nm23-H2WT, pcDNA3-Myc-Nm23-H2H118C, and pcDNA3-Nm23-H2- Kpn were prepared by PCR using the pGad424-Nm23-H2 plasmid (isolated by yeast two-hybrid screening) and the following primers, respectively, NM23F (5 -GAGGAATTCATGGCCAACCTGGAGCGCACC-3 ), NM23R (5 -GCGCTCGAGTCACAGATCCTCTTCAGAGATGAGTTTCTGTTCTTCATAGACCCAGTCATG-3 ), Nm23H118CF (5 -GGCAGGAACATCATTTGTGGCAGTGATTCAG-3 ), NM23H118CR (5 -CTGAATCACTGCCACAAAGATGTTCCTGCC-3 ), NM23 KpnF (5 -CTGAATCACTGCATGAATGATGGTCTCCCCAAGCATCACTCGGCC-3 ), and NM23 KpnR (5 -GGCCGAGTGATGCTTGGGGAGACCATCATTCATGGCAGTGATTCAG-3 ). ..

    Clone Assay:

    Article Title: Virion-Wide Protein Interactions of Kaposi's Sarcoma-Associated Herpesvirus
    Article Snippet: .. The cDNAs for 11 tegument proteins or fragments were cloned into a Clontech Matchmaker system two-bait vector pAS2-1, which allows expression of a fusion protein of the DNA binding domain of yeast GAL4 protein and a tegument protein. cDNAs of all KSHV virion proteins (capsid, tegument, and envelope glycoproteins) were cloned into Matchmaker prey vector pACT2, generating the prey array listed in Table 1. ..

    Expressing:

    Article Title: Virion-Wide Protein Interactions of Kaposi's Sarcoma-Associated Herpesvirus
    Article Snippet: .. The cDNAs for 11 tegument proteins or fragments were cloned into a Clontech Matchmaker system two-bait vector pAS2-1, which allows expression of a fusion protein of the DNA binding domain of yeast GAL4 protein and a tegument protein. cDNAs of all KSHV virion proteins (capsid, tegument, and envelope glycoproteins) were cloned into Matchmaker prey vector pACT2, generating the prey array listed in Table 1. ..

    Binding Assay:

    Article Title: Virion-Wide Protein Interactions of Kaposi's Sarcoma-Associated Herpesvirus
    Article Snippet: .. The cDNAs for 11 tegument proteins or fragments were cloned into a Clontech Matchmaker system two-bait vector pAS2-1, which allows expression of a fusion protein of the DNA binding domain of yeast GAL4 protein and a tegument protein. cDNAs of all KSHV virion proteins (capsid, tegument, and envelope glycoproteins) were cloned into Matchmaker prey vector pACT2, generating the prey array listed in Table 1. ..



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    Image Search Results


    Identification of I-2 domains required for yeast two-hybrid (Y2H) interaction with I2I-1 and I2I-2. (A) Y2H analyses show interactions of I2I-1 or I2I-2 with full-length I-2 as bait, but not with Rx and Mi-1.2. I2I-1 and I2I-2 interact with the CC-NB-ARC bait of I-2 (amino acids 1–520), but not with bait constructs containing the CC-NB-ARC domains of I-2C1 (amino acids 1–526), Rx (amino acids 1–426), or Mi-1.2 (amino acids 161–896). The baits and preys did not show autoactivity when co-expressed with the pACT2 and pAS2-1 empty vectors. The test for activation of the HIS3 marker is shown after 10 days of growth (left panel). Expression of the bait proteins was confirmed by Western blot (WB) analyses using α-Gal4BD antibody detection on total yeast protein extracts (right panel). (B) Y2H analyses of I2I-1 and I2I-2 with various I-2 baits: I-2FL (full-length: amino acids 1–1266), I-2N+ (amino acids 1–643), I-2N (amino acids 1–520), I-2N ΔMHD (amino acids 1–520, with deletion at amino acids 484–496), and I-2CC (amino acids 1–170). The different I-2 (sub)domains are indicated and coloured: orange, coiled-coil (CC); red, nucleotide-binding (NB); purple, Apaf-1, R proteins, and Ced-4 (ARC) 1; blue, ARC2; and green, leucine-rich repeat (LRR). The test for activation of the ADE2 marker is shown after 5 days of growth; the other two markers ( HIS3, LacZ ) gave identical results. +, activation of all three selectable markers; –, no activation of these markers. The smallest part of I-2 enabling interaction with I2I-1 is the CC domain and for I2I-2 the CC-NB-ARC domain.

    Journal: Journal of Experimental Botany

    Article Title: Protein–protein interactions as a proxy to monitor conformational changes and activation states of the tomato resistance protein I-2

    doi: 10.1093/jxb/ers021

    Figure Lengend Snippet: Identification of I-2 domains required for yeast two-hybrid (Y2H) interaction with I2I-1 and I2I-2. (A) Y2H analyses show interactions of I2I-1 or I2I-2 with full-length I-2 as bait, but not with Rx and Mi-1.2. I2I-1 and I2I-2 interact with the CC-NB-ARC bait of I-2 (amino acids 1–520), but not with bait constructs containing the CC-NB-ARC domains of I-2C1 (amino acids 1–526), Rx (amino acids 1–426), or Mi-1.2 (amino acids 161–896). The baits and preys did not show autoactivity when co-expressed with the pACT2 and pAS2-1 empty vectors. The test for activation of the HIS3 marker is shown after 10 days of growth (left panel). Expression of the bait proteins was confirmed by Western blot (WB) analyses using α-Gal4BD antibody detection on total yeast protein extracts (right panel). (B) Y2H analyses of I2I-1 and I2I-2 with various I-2 baits: I-2FL (full-length: amino acids 1–1266), I-2N+ (amino acids 1–643), I-2N (amino acids 1–520), I-2N ΔMHD (amino acids 1–520, with deletion at amino acids 484–496), and I-2CC (amino acids 1–170). The different I-2 (sub)domains are indicated and coloured: orange, coiled-coil (CC); red, nucleotide-binding (NB); purple, Apaf-1, R proteins, and Ced-4 (ARC) 1; blue, ARC2; and green, leucine-rich repeat (LRR). The test for activation of the ADE2 marker is shown after 5 days of growth; the other two markers ( HIS3, LacZ ) gave identical results. +, activation of all three selectable markers; –, no activation of these markers. The smallest part of I-2 enabling interaction with I2I-1 is the CC domain and for I2I-2 the CC-NB-ARC domain.

    Article Snippet: The bait used for the library screen was constructed by subcloning the Nco I/ Sac I fragment of I-2 FL in the yeast two-hybrid (Y2H) bait vector pAS2-1 (Clontech Laboratories, Palo Alto, CA, USA), resulting in bait CC-NB-ARC-LRR1-12 (amino acids 1–872).

    Techniques: Construct, Activation Assay, Marker, Expressing, Western Blot, Binding Assay