title: Cloning, expression, and preliminary characterization of the dysferlin tegument protein in Schistosoma japonicum.
abstract: The schistosomal tegument is a dynamic host-interactive layer. Proteins exposed to the host on the tegumental surface are important for completion of the parasitic lifecycle. Dysferlin is a member of the ferlin family and is involved in plasma membrane repair. Based on the results of a proteomics study of tegument surface proteins of Schistosoma japonicum in our laboratory, dysferlin was identified as a tegumental protein of S. japonicum. The gene encoding S. japonicum dysferlin (SjDF), which codes for several Ca(2+) binding sites, was cloned, expressed in Escherichia coli, and characterized. Western blot analysis revealed that recombinant SjDF had good immunogenicity. Real-time RT-PCR analysis showed that SjDF was upregulated mainly in adult worms and the transcription level in 42-day-old female worms was significantly higher than that in males. Immunofluorescence analysis revealed that SjDF was mainly distributed in the tegument at various developmental stages. Experimental mice were treated with praziquantel and at 35days post-infection, we noted that damage to the tegument and subtegument worsened and did not recover at 36h post-treatment in the high-dose group and was accompanied by downregulation of SjDF mRNA, while the damage was less severe and recovered by this time in the low-dose group, and accompanied by upregulation of SjDF. Our results suggested that SjDF is a tegumental protein that may be important in schistosomal development and may participate in the repair process in muscle and tegument, and could present a viable vaccine candidate for schistosomiasis.
authors.affiliation: Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai 200241, PR China.
authors.forename: Yanian
authors.initials: Y
authors.lastname: Xiong
authors.forename: Dezhou
authors.initials: D
authors.lastname: Ai
authors.forename: Peipei
authors.initials: P
authors.lastname: Meng
authors.forename: Meimei
authors.initials: M
authors.lastname: Wei
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
authors.forename: Lini
authors.initials: L
authors.lastname: Huang
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Fu
authors.forename: Yaojun
authors.initials: Y
authors.lastname: Shi
authors.forename: Jiaojiao
authors.initials: J
authors.lastname: Lin
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D018509:DNA, Helminth / Q000737:chemistry / Q000235:genetics; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D005260:Female; D005786:Gene Expression Regulation; D015801:Helminth Proteins / Q000235:genetics* / Q000378:metabolism; D008168:Lung / Q000469:parasitology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D017422:Sequence Analysis, DNA; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000909:Antibodies, Helminth; D018076:DNA, Complementary; D018509:DNA, Helminth; D015801:Helminth Proteins; D012333:RNA, Messenger; D011994:Recombinant Proteins
keywords: Dysferlin; Plasma membrane repair; Schistosoma japonicum