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Expression and characterization of a phospholipid hydroperoxide glutathione peroxidase gene in Schistosoma japonicum

Published online by Cambridge University Press:  18 August 2015

YING ZHANG
Affiliation:
Department of Medical Genetics, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China
YUAN HE
Affiliation:
Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China
LI HE
Affiliation:
Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China
HONGYING ZONG
Affiliation:
Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China
GUOBIN CAI*
Affiliation:
Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China
*
*Corresponding author. Department of Medical Parasitology, Wuhan University School of Basic Medical Sciences, Wuhan 430071, China. E-mail: [email protected]

Summary

Phospholipid hydroperoxide glutathione peroxidase (PHGPx, GPx4) is a major antioxidant enzyme, which plays unique roles in the protection of cells against oxidative stress by catalysing reduction of lipid hydroperoxides. We isolated and characterized a full-length cDNA sequence encoding GPx gene from a blood fluke, Schistosoma japonicum (designated SjGPx), which contained an in-frame TGA codon for selenocysteine (Sec) and a concurrent Sec insertion sequence in its 3′-untranslated region. Protein encoded by SjGPx demonstrated a primary structure characteristic to the PHGPx family, including preservation of catalytic domains and absence of the subunit interaction domains. Semi-quantitative reverse transcription PCR and Western blotting showed that the SjGPx was mainly expressed in the female adults and eggs. RNA interference approach was employed to investigate the effects of knockdown of SjGPx. SjGPx expression level was significantly reduced on the 5th day post-RNAi. Significantly reduction in GPx enzyme activities, as well as obvious changes in morphology of intrauterine eggs followed the reduction in SjGPx transcript level. We observed a 63·04% reduction in GPx activity and the eggs severely deformed. Our results revealed that SjGPx protein might be involved in the provision of enzyme activity during egg production.

Type
Research Article
Copyright
Copyright © Cambridge University Press 2015 

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