Related Experiment Video
Updated: Aug 13, 2026
High-Throughput Expression and Purification of Human Solute Carriers for Structural and Biochemical Studies
Published on: September 29, 2023
Molecular cloning and characterization of cDNA encoding a novel cytosolic glutathione transferase from Clonorchis
Zhongluan Wu1, De Wu, Xuchu Hu
1Department of Parasitology, School of Pre-Clinical Medicine, SUN Yat-Sen University, Guangzhou, 510089, China.
Abstract:
Glutathione transferases (GSTs) are a group of multifunction isoenzymes coded by many genes. A cDNA encoding a novel cytosolic GST enzyme was cloned from a Clonorchis sinensis (Cs) adult worm cDNA library by large-scale sequencing. This new cDNA contains 786 bp with a putative open reading frame of 212 amino acids. The deduced amino acid sequence exhibits 61% identity to C. sinensis cytosolic 28-kDa GST. Recombinant CsGST was overexpressed in Escherichia coli BL21(DE3) and was purified by Ni-NTA Agarose. Enzymatic assays showed that the recombinant CsGST had a high activity of 22.7 U mg(-1). The average K (m) of the CsGST for 1-chloro-2,4-dinitrobenzene is 111 microM. Cibacron blue F3GA and albendazole inhibited the CsGST activity with respective average IC(50) of 1.1 and 247.1 microM. It provides a model for the structure and physiological function analysis on CsGST. The nucleotide sequence reported in this paper has been submitted to the GenBank Database with accession number DQ179264.
