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matchmaker pre transformed hela cdna library  (TaKaRa)


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    TaKaRa matchmaker pre transformed hela cdna library
    (A) A yeast-two hybrid screen, using a <t>HeLa</t> <t>cDNA</t> library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.
    Matchmaker Pre Transformed Hela Cdna Library, supplied by TaKaRa, used in various techniques. Bioz Stars score: 90/100, based on 113 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/matchmaker+pre+transformed+hela+cdna+library/pmc05176192-220-1-6?v=TaKaRa
    Average 90 stars, based on 113 article reviews
    matchmaker pre transformed hela cdna library - by Bioz Stars, 2026-08
    90/100 stars

    Images

    1) Product Images from "The Effector Cig57 Hijacks FCHO-Mediated Vesicular Trafficking to Facilitate Intracellular Replication of Coxiella burnetii"

    Article Title: The Effector Cig57 Hijacks FCHO-Mediated Vesicular Trafficking to Facilitate Intracellular Replication of Coxiella burnetii

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1006101

    (A) A yeast-two hybrid screen, using a HeLa cDNA library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.
    Figure Legend Snippet: (A) A yeast-two hybrid screen, using a HeLa cDNA library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.

    Techniques Used: Two Hybrid Screening, cDNA Library Assay, Binding Assay, Plasmid Preparation, Pull Down Assay, Infection, Expressing, Transfection, Construct, Staining, Stable Transfection



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    (A) A yeast-two hybrid screen, using a <t>HeLa</t> <t>cDNA</t> library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.
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    Image Search Results


    (A) A yeast-two hybrid screen, using a HeLa cDNA library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.

    Journal: PLoS Pathogens

    Article Title: The Effector Cig57 Hijacks FCHO-Mediated Vesicular Trafficking to Facilitate Intracellular Replication of Coxiella burnetii

    doi: 10.1371/journal.ppat.1006101

    Figure Lengend Snippet: (A) A yeast-two hybrid screen, using a HeLa cDNA library, revealed FCHO2 as a binding partner of Cig57. Growth on double dropout (DDO) plates for all transformations indicates plasmid retention and yeast viability. Growth on quadruple dropout (QDO) plates indicates the interaction between FCHO2 and Cig57 (segment 2) and the lack of growth in segment 1 and 3 of this plate indicates no interaction with the empty vectors present. (B) A Pull-down assay verified the interaction between Cig57 and GFP-FCHO2 during Coxiella infection of HEK 293T cells. Cells were infected with wild-type (WT) Coxiella , or Coxiella cig57 ::Tn strain expressing 3xFLAG-Cig57 from a plasmid, and transfected with GFP constructs (GFP or GFP-FCHO2). GFP proteins were pulled out using GFP-Trap beads. The presence of 3xFLAG-Cig57 in the pull down lysates indicates interaction between GFP-FCHO2 and Cig57. Results are shown for one repeat of the experiment. (C) Representative images showing HeLa cells transfected to express mCherry or mCherry-Cig57 (red) during infection with wild-type C . burnetii (green). Nuclei are stained in blue with DAPI. Scale bar = 10 μm. (D) HeLa cells stably expressing GFP-FCHO2 were transfected with pmCherry-Cig57 and nuclei stained with DAPI (blue). Scale bars represent 10 μm and the CCV is denoted by an asterisk.

    Article Snippet: The Matchmaker pre-transformed HeLa cDNA library (Clontech) was mated with S . cerevisiae carrying pGBKT7- cig57 according to manufacturer’s protocols (Clontech PT3183-1 manual).

    Techniques: Two Hybrid Screening, cDNA Library Assay, Binding Assay, Plasmid Preparation, Pull Down Assay, Infection, Expressing, Transfection, Construct, Staining, Stable Transfection