Protocol detail

Purification of His- and GST-tagged TatD-like DNase

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The genes encoding the His- and GST-tagged TatD-like DNase of P. berghei (PBANKA_0201800) were cloned into the pET-28a and pGEX-4T-1 vectors, respectively (Invitrogen), and expressed in Escherichia coli BL21(DE3), as described in our earlier study (Chang et al., 2016 (link)). His- and GST-tagged recombinant proteins were purified using the His GraviTrap^TM system (GE Healthcare) and the Glutathione Sepharose^TM 4B system (GE Healthcare), respectively, according to the manufacturer’s instructions. Purified proteins were analyzed with SDS-PAGE and Western blots before further experiments.

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Wang W., Liu F., Jiang N., Lu H., Yang N., Feng Y., Sang X., Cao Y, & Chen Q. (2018). Plasmodium TatD-Like DNase Antibodies Blocked Parasite Development in the Mosquito Gut. Frontiers in Microbiology, 9, 1023.

Publication 2018

pET-28a pGEX-4T-1 His GraviTrapTM system Glutathione SepharoseTM 4B system

Dnase Escherichia coli Genes Glutathione Proteins Recombinant proteins Sds page Vectors

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Variable analysis

independent variables

Cloning of the genes encoding the His- and GST-tagged TatD-like DNase of P. berghei (PBANKA_0201800) into the pET-28a and pGEX-4T-1 vectors, respectively
Expression of the His- and GST-tagged recombinant proteins in Escherichia coli BL21(DE3)

dependent variables

Purification of the His- and GST-tagged recombinant proteins using the His GraviTrap™ system and the Glutathione Sepharose™ 4B system, respectively
Analysis of the purified proteins using SDS-PAGE and Western blots

control variables

Manufacturer's instructions for the His GraviTrap™ system and the Glutathione Sepharose™ 4B system

controls

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

Annotations

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