Recombinant Viral Capsid Protein Production

The cDNAs encoding the capsid P domain were cloned into the expression vector pGEX-4T-1 (Amersham Biosciences, Piscataway, NJ, United States) between BamHI and NotI sites. After sequence confirmation, P proteins were expressed in Escherichia coli following previously described procedures (Tan et al., 2008 (link)). Briefly, The BL21 cultures were initiated by 0.5 mM IPTG (isopropyl-β-D-thiogalactopyranoside) at a temperature of 22°C overnight. The recombinant P domain-GST fusion proteins were purified using Glutathione Sepharose 4 Fast Flow resin (GE Healthcare Life Sciences, NJ, United States). The GST was removed from the P proteins by thrombin (GE Healthcare Life Sciences, Princeton, NJ, United States) cleavage on beads at 22°C overnight.

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Xie D., Chen J., Yu J., Pei F., Koroma M.M., Wang L., Qiu M., Hou Y., Yu D., Zhang X.F, & Dai Y.C. (2020). Characterization of Antigenic Relatedness Among GI Norovirus Genotypes Using Serum Samples From Norovirus-Infected Patients and Mouse Sera. Frontiers in Microbiology, 11, 607723.

Publication 2020

pGEX-4T-1 Glutathione Sepharose 4 Fast Flow resin thrombin

Capsid Cdnas Cleavage Escherichia coli Glutathione Iptg P proteins Recombinant fusion proteins Resin Sepharose Thrombin Vector

Corresponding Organization : Southern Medical University

Other organizations : Nanfang Hospital, Guangzhou Center for Disease Control and Prevention

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Variable analysis

independent variables

IPTG concentration (0.5 mM)

dependent variables

Expression of recombinant P domain-GST fusion proteins

control variables

Temperature (22°C)
Overnight incubation duration

controls

Positive control: Not mentioned
Negative control: Not mentioned

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