Recombinant Fn-Dps Protein Production

Fn-Dps was obtained as we mentioned in our primary study^{17 (link)}. In brief, Fn-Dps gene was amplified by polymerase chain reaction (PCR) and Fn (ATCC 25586) DNA as templates. The recombinant plasmids of pET28a-Fn-Dps were transformed into E. coli BL21 (DE3). The expression of the recombinant Fn-Dps was induced with 0.8 mM isopropyl β-d-1-thiogalactopyranoside (IPTG) for continuous 16 h cultivation at 25 °C. The endotoxin in Fn-Dps was removed using the Endotoxin Removal Kit (GenScript, Nanjing, China) according to the manufacturer’s instructions.

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Li Y., Xing S., Chen F., Li Q., Dou S., Huang Y., An J., Liu W, & Zhang G. (2023). Intracellular Fusobacterium nucleatum infection attenuates antitumor immunity in esophageal squamous cell carcinoma. Nature Communications, 14, 5788.

Publication 2023

Endotoxin Removal Kit

E coli Endotoxin Gene Plasmids Polymerase chain reaction

Corresponding Organization : Third Affiliated Hospital of Sun Yat-sen University

Other organizations : Sun Yat-sen University Cancer Center

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Variable analysis

independent variables

Expression of the recombinant Fn-Dps was induced with 0.8 mM isopropyl β-d-1-thiogalactopyranoside (IPTG)

dependent variables

Expression level of the recombinant Fn-Dps

control variables

Continuous 16 h cultivation at 25 °C
Fn (ATCC 25586) DNA as templates for PCR amplification of Fn-Dps gene
Transformation of recombinant plasmids of pET28a-Fn-Dps into E. coli BL21 (DE3)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

Annotations

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