title: Molecular characterization of Clonorchis sinensis tetraspanin 2 extracellular loop 2.
abstract: In the course of examining EST of the liver fluke Clonorchis sinensis, a cDNA encoding the protein similar to tetraspanin 2 (TSP-2) of blood fluke schistosome was identified as CsTSP2. TSPs are a family of eukaryotic cell membrane-spanning proteins thought to anchor multiple proteins to one area of the cell membrane. Multiple sequence alignment of CsTSP2 revealed over 40% of identities with those of schistosomes. The CCC, PXSC, and CG motives characteristic to extracellular loop 2 (EC2) region of TSP-2 were well conserved. PCR-amplified EC2 of CsTSP2 (CsTSP2EC2) was subcloned into pEXP5 NT/TOPO bacterial expression plasmid vector. Recombinant CsTSP2EC2 protein (rCsTSP2EC2) fused with 6X His tag was overexpressed bacterially and purified by using Ni-NTA affinity chromatography under denaturing condition. The purified rCsTSP2EC2 was recognized specifically by the sera from human infected with C. sinensis. Considering biological role and specific immunity of TSPs, rCsTSP2EC2 might be a probable vaccine candidate against clonorchiasis. Protective effects of immunizing rCsTSP2EC2 should be further studied.
authors.affiliation: Department of Environmental Medical Biology and Institute of Tropical Medicine, Yonsei University College of Medicine, Seoul 120-752, South Korea.
authors.forename: Tae Yun
authors.initials: TY
authors.lastname: Kim
authors.forename: Eun Joo
authors.initials: EJ
authors.lastname: Chung
authors.forename: Woon-Mok
authors.initials: WM
authors.lastname: Sohn
authors.forename: Sung-Hong
authors.initials: SH
authors.lastname: Hong
authors.forename: Tai-Soon
authors.initials: TS
authors.lastname: Yong
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D003003:Clonorchiasis / Q000276:immunology; D003004:Clonorchis sinensis / Q000235:genetics*; D017124:Conserved Sequence; D015870:Gene Expression; D015801:Helminth Proteins / Q000235:genetics*; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D059470:Tetraspanins / Q000235:genetics*
chemical_list: D000909:Antibodies, Helminth; D015801:Helminth Proteins; D011993:Recombinant Fusion Proteins; D059470:Tetraspanins