title: Molecular cloning, expression, and immunolocalization of the NAD(+)-dependent glycerol 3-phosphate dehydrogenase (GPD) from Clonorchis sinensis.
abstract: Glycerol 3-phosphate dehydrogenase (GPD) plays an important role in the energy metabolism and nutrition metabolism. In order to know about the biological functions of GPD of Clonorchis sinensis (C. sinensis), we identified a complete gene coding GPD from C. sinensis metacercaria cDNA library. This novel cDNA sequence contains 1,056 bp with a putative open reading frame of 351 amino acids and shares 74% identity with GPD from Schistosoma mansoni. Recombinant CsGPD was expressed and purified from Escherichia coli BL21 (DE3). Western blot analysis displayed that recombinant CsGPD can be recognized by anti-CsGPD serum and C. sinensis-infected serum. RT-PCR and immunolocalization analysis confirmed that GPD expressed both at the stage of adult worm and metacercaria of C. sinensis and immunolocated at the tegument of adult worm, tegument and tegumentary cells of metacercaria. Our current study has paved the way for the further researches about the biological functions involved in the growth of C. sinensis.
authors.affiliation: Department of Parasitology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, People's Republic of China.
authors.forename: Yongxiu
authors.initials: Y
authors.lastname: Fan
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Wang
authors.forename: Chuanhuan
authors.initials: C
authors.lastname: Deng
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.forename: Lexun
authors.initials: L
authors.lastname: Wang
authors.forename: Wenjun
authors.initials: W
authors.lastname: Chen
authors.forename: Chi
authors.initials: C
authors.lastname: Liang
authors.forename: Xuerong
authors.initials: X
authors.lastname: Li
authors.forename: Zhongdao
authors.initials: Z
authors.lastname: Wu
authors.forename: Xinbing
authors.initials: X
authors.lastname: Yu
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D003001:Cloning, Molecular; D003004:Clonorchis sinensis / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+) / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D016366:Open Reading Frames; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
chemical_list: D000909:Antibodies, Helminth; D011994:Recombinant Proteins; D050536:Glycerol-3-Phosphate Dehydrogenase (NAD+)