| Echinococcosis is a serious zoonotic parasitic disease caused by the larvae of Echinococcus granulosus parasitizing the visceral organs of humans and animals.The disease is distributed worldwide,and China is one of the high-incidence areas.At present,there are reports of primary echinococcosis in 23 provinces and 368 counties of China,with the highest incidence in Northwest Sichuan,Xinjiang,Qinghai and Tibet pastoral areas.It has caused serious economic losses to animal husbandry in China,and also brought many environmental health and safety problems to human beings.In this study,we amplified phosphoglycerate kinase(Eg-PGK)and ubiquitin-binding enzyme D2(Eg-E2D2)genes of Echinococcus granulosus,and analyzed their basic characteristics,providing important reference data for the study of Echinococcus granulosus related proteins.In addition,the recombinant proteins of fatty acid binding protein with A31 protein(EgFABP-EgA31)and thioredoxin peroxidase with tropomyosin(EgTPx-EgTrp)were cloned and expressed.The protective effect of the recombinant proteins was evaluated after immunizing dogs.1.Cloning,expression and characterization of phosphoglycerate kinase gene from Echinococcus granulosusIn this study,we cloned,expressed and identified Eg-PGK.The result showed that the sizes of Eg-PGK was 1248 bp.Western blotting showed that rEg-PGK had reactionogenicity.The indirect fluorescence immunoassay showed that Eg-PGK was mainly distributed in the tegument and hooks of protoscoleces,tegument,parenchyma and eggs of adults,as well as in fertile and infertile cysts.Quantitative PCR analysis showed that the transcription level of Eg-PGK was no significant difference between protoscolece stage and 18-day-old larvae stage(p>0.05).In conclusion,Eg-PGK may plays an important role in the growth and development of Echinococcus granulosus,and participates in tegument repair and anti-host immunity.This study provides basic data for future research on Eg-PGK.2.Cloning,expression and characterization of ubiquitin-conjugating enzyme D2 gene from Echinococcus granulosusIn this study,we cloned,expressed and identified Eg-E2D2.The result showed that the sizes of Eg-E2D2 was 1048 bp.Western blotting showed that rEg-E2D2 had reactionogenicity.The indirect fluorescence immunoassay showed that Eg-E2D2 was distributed in the tegument and hooks of protoscoleces,parenchyma of adults,as well as infertile and infertile cysts.Quantitative PCR analysis showed that the transcription level of Eg-E2D2 was no significant difference between protoscolece stage and 18-day-old larvae stage(p>0.05).In conclusion,Eg-E2D2 may plays an important role in the growth and development of Echinococcus granulosus,and is essential to the worm.This study provides basic data for future research on Eg-E2D2.3.Evaluation of immunoprotective effect of two fusion proteins(EgFABP-EgA31 and EgTPx-EgTrp)of Echinococcus granulosus on dogsIn this study,we cloned and expressed recombinant proteins of EgFABP-EgA31 and EgTPx-EgTrp.rEgFABP-EgA31 and rEgTPx-EgTrp protein were immunized in each group of Beagle dogs.After vaccination,each Beagle dog was orally challenge-infected protoscoleces of Echinococcus granulosus.The antibody level was measured,and the number and length and width of the adults were measured after the Beagle dogs were sacrificed.The results showed that the average number of worms in EgFABP-EgA31 and EgTPx-EgTrp groups was less than that in the control group(p < 0.05),the worm reduction rates were 67.00% and 55.43% respectively,and the length and width of the adults in the immune group were less than those in the control group.Antibody level showed that the immune group had higher antibody level,but the increase and decrease of antibody level in EgFABP-EgA31 group was more stable than that in EgTPx-EgTrp group.EgFABP-EgA31 group and EgTPx-EgTrp group had protective effect on dogs,and EgFABP-EgA31 group had better effect than EgTPx-EgTrp group. |
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