The gene fragment coding for TgTCP-1 was cloned into the pGEX-4T-1 and pET-28a vectors, respectively (Invitrogen, Carlsbad, CA, United States), and the recombinant plasmids were transformed into E. coli BL21 (DE3) for protein expression, respectively. The GST- and His-tagged fusion TgTCP-1 proteins were purified using the Glutathione SepharoseTM 4B system (GE Healthcare) and the His GraviTrapTM system (GE Healthcare), respectively, according to the manufacturer’s instructions. The purified proteins were verified by SDS-PAGE and Western blotting.
His gravitrap system
Manufactured by GE Healthcare
The GraviTrapTM system is a lab equipment product developed by GE Healthcare. It is designed to facilitate the separation and isolation of particles or molecules based on their density or gravitational force. The core function of the GraviTrapTM system is to provide a controlled and efficient method for density-based separation and purification of samples.
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2 protocols using his gravitrap system
1
Cloning and Purification of TgTCP-1
2
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 GraviTrapTM system (GE Healthcare) and the Glutathione SepharoseTM 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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