Pcold gst plasmid

Manufactured by Takara Bio

The PCold GST plasmid is a bacterial expression vector designed for the production of recombinant proteins fused with glutathione S-transferase (GST) tag. It can be used to express target proteins in Escherichia coli under cold-shock induction conditions.

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Lab products found in correlation

Pcold tf plasmid, by Takara Bio (3 mentions) Gst protein interaction pull down kit, by Thermo Fisher Scientific (2 mentions) Bl21 competent cells, by Vazyme (2 mentions)

3 protocols using pcold gst plasmid

PEDV Protein Interaction Analysis

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PEDV N, MARCH8, NDP52, and RALY gene sequences were cloned into either pCold GST plasmid (Clontech Laboratories, Inc; catalog no.: 3372) or pCold TF plasmid (Clontech Laboratories, Inc; catalog no.: 3365) and expressed in BL21-competent cells (Vazyme Biotech; catalog no.: C504-03). The GST Protein Interaction Pull-Down Kit (Thermo; catalog no.: 21516) was used to analyze protein interactions, according to the manufacturer’s instructions. The proteins were eluted with reduced glutathione and analyzed by Western blotting.

Qin W., Kong N., Zhang Y., Dong S., Zhai H., Zhai X., Yang X., Ye C., Ye M., Liu C., Yu L., Zheng H., Yu H., Zhang W., Tong G., Lan D., Tong W, & Shan T. (2022). Nuclear ribonucleoprotein RALY targets virus nucleocapsid protein and induces autophagy to restrict porcine epidemic diarrhea virus replication. The Journal of Biological Chemistry, 298(8), 102190.

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PEDV N-PABPC4 Interaction Assay

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PEDV N was cloned into pCold GST plasmid (Clontech), and PABPC4 was cloned into pCold TF plasmid (Clontech). Recombinant proteins were expressed in BL21 cells (Vazyme), following which cells were lysed using the lysis buffer provided in the GST protein interaction pulldown kit (Thermo) and incubated on ice for 30 min. After centrifugation, equal amounts of GST and GST-N supernatants were mixed with PABPC4. GST pulldown assays were performed using the GST protein interaction pulldown kit according to the manufacturer's protocol. The proteins were eluted with reduced glutathione and analyzed by Western blotting.

Jiao Y., Kong N., Wang H., Sun D., Dong S., Chen X., Zheng H., Tong W., Yu H., Yu L., Huang Y., Wang H., Sui B., Zhao L., Liao Y., Zhang W., Tong G, & Shan T. (2021). PABPC4 Broadly Inhibits Coronavirus Replication by Degrading Nucleocapsid Protein through Selective Autophagy. Microbiology Spectrum, 9(2), e00908-21.

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Cloning and Protein Interaction Analysis

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The full-length PEDV N, MARCH8, NDP52, and PTBP1 genes were cloned into the pCold GST plasmid (3372, Clontech Laboratories, Inc) or pCold TF plasmid (3365, Clontech Laboratories, Inc). The plasmids were expressed in BL21 competent cells (C504–03, Vazyme Biotech Co, Ltd). The GST Tag Protein Interaction Pull-Down Kit (21516, Thermo Fisher Scientific Pierce) was used for protein interaction analysis in line with specific protocols. After elution, Western blot assay was performed for protein analysis.

Qin W., Kong N., Zhang Y., Wang C., Dong S., Zhai H., Zhai X., Yang X., Ye C., Ye M., Tong W., Liu C., Yu L., Zheng H., Yu H., Zhang W., Lan D., Tong G, & Shan T. (2023). PTBP1 suppresses porcine epidemic diarrhea virus replication via inducing protein degradation and IFN production. The Journal of Biological Chemistry, 299(8), 104987.

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