title: Identification, expression and phylogenetic analysis of EgG1Y162 from Echinococcus granulosus.
abstract: OBJECTIVE
  This study was to clone, identify and analyze the characteristics of egG1Y162 gene from Echinococcus granulosus.
  
  
  METHODS
  Genomic DNA and total RNAs were extracted from four different developmental stages of protoscolex, germinal layer, adult and egg of Echinococcus granulosus, respectively. Fluorescent quantitative PCR was used for analyzing the expression of egG1Y162 gene. Prokaryotic expression plasmid of pET41a-EgG1Y162 was constructed to express recombinant His-EgG1Y162 antigen. Western blot analysis was performed to detect antigenicity of EgG1Y162 antigen. Gene sequence, amino acid alignment and phylogenetic tree of EgG1Y162 were analyzed by BLAST, online Spidey and MEGA4 software, respectively.
  
  
  RESULTS
  EgG1Y162 gene was expressed in four developmental stages of Echinococcus granulosus. And, egG1Y162 gene expression was the highest in the adult stage, with the relative value of 19.526, significantly higher than other three stages. Additionally, Western blot analysis revealed that EgG1Y162 recombinant protein had good reaction with serum samples from Echinococcus granulosus infected human and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%.
  
  
  CONCLUSIONS
  Our study identified EgG1Y162 antigen in Echinococcus granulosus for the first time. EgG1Y162 antigen had a high similarity with EmY162 antigen, with the genetic differences mainly existing in the intron region. And, EgG1Y162 recombinant protein showed good antigenicity.
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Fengbo
authors.initials: F
authors.lastname: Zhang
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Xiumin
authors.initials: X
authors.lastname: Ma
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Yuejie
authors.initials: Y
authors.lastname: Zhu
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Hongying
authors.initials: H
authors.lastname: Wang
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Xianfei
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Immunology, Xinjiang Medical University Urumqi 830011, Xinjiang, P.R. China.
authors.forename: Min
authors.initials: M
authors.lastname: Zhu
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Haimei
authors.initials: H
authors.lastname: Ma
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Wen
authors.affiliation: Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University 251 Xining Road, Xining 810000, Qinghai, China.
authors.forename: Haining
authors.initials: H
authors.lastname: Fan
authors.affiliation: State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China ; Department of Immunology, Xinjiang Medical University Urumqi 830011, Xinjiang, P.R. China.
authors.forename: Jianbing
authors.initials: J
authors.lastname: Ding
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000947:Antigens, Helminth / Q000097:blood / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015153:Blotting, Western; D003001:Cloning, Molecular; D030541:Databases, Genetic; D004285:Dogs; D004443:Echinococcosis / Q000097:blood / Q000469:parasitology*; D048209:Echinococcus granulosus / Q000235:genetics / Q000254:growth & development / Q000276:immunology*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D015800:Protozoan Proteins / Q000097:blood / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000276:immunology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
chemical_list: D000913:Antibodies, Protozoan; D000947:Antigens, Helminth; D015800:Protozoan Proteins; D011994:Recombinant Proteins
keywords: EgG1Y162 antigen; fluorescent quantitative PCR; phylogenetic tree; prokaryotic expression