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Molecular characterization and tissue localization of glutathione S-transferase from adult Ancylostoma ceylanicum

  • J.X. Hang (a1), L. He (a1), A.M.I. Abuzeid (a1), Y. Huang (a1), Y.Q. Liu (a1), X.X. Yan (a1), Q. Zhao (a1), X. Li (a1), J.M. Liu (a1) and G.Q. Li (a1)...

Abstract

Glutathione S-transferases (GSTs) are a detoxifying enzyme family that is essential for parasite blood-feeding and survival, and represent potential targets for hookworm vaccine development. Multiple GST-encoding complementary DNAs (cDNAs) have been cloned from Ancylostoma caninum and Necator americanus, but there are no reports about the cloning of this enzyme from Ancylostoma ceylanicum, the animal-derived zoonotic hookworm. To study the molecular nature and tissue localization of GST of A. ceylanicum (Ace-GST), we designed primers based on the GST gene sequence of A. ceylanicum in GenBank, amplified the Ace-GST cDNA by reverse transcription polymerase chain reaction, and analysed its homology and genetic evolution relationship. The amplified product was cloned into the pET-32a vector and transformed into Escherichia coli BL21 (DE3) for expression. To prepare anti-GST polyclonal antibodies, the recombinant protein was purified and used to immunize Kunming mice. The level of immunoglobulin G (IgG) antibody in the serum of immunized mice was detected by indirect enzyme-linked immunosorbent assay, and the Ace-GST localization in adult worm was determined using the immunofluorescence method. The results showed that the full-length cDNA encoding Ace-GST was 468 bp, which had the highest homology with Ac-GST-1 (60.1%) and clustered into one branch (v-class) with Ac-GST-1 and Na-GST-1 in a phylogenetic tree. Mice immunized with recombinant Ace-GST showed specific IgG antibody response. Immunolocalization revealed that natural Ace-GST is mainly located in the epidermis, muscle and intestine of the adult. These results may lay a foundation for further studies on the biological function of Ace-GST.

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Corresponding author

Author for correspondence: G.Q. Li, E-mail: gqli@scau.edu.cn

References

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Keywords

Molecular characterization and tissue localization of glutathione S-transferase from adult Ancylostoma ceylanicum

  • J.X. Hang (a1), L. He (a1), A.M.I. Abuzeid (a1), Y. Huang (a1), Y.Q. Liu (a1), X.X. Yan (a1), Q. Zhao (a1), X. Li (a1), J.M. Liu (a1) and G.Q. Li (a1)...

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