Fengbo Zhang1, Xiumin Ma1, Yuejie Zhu1, Hongying Wang1, Xianfei Liu1, Min Zhu2, Haimei Ma1, Hao Wen1, Haining Fan3, Jianbing Ding4. 1. State Key Laboratory Incubation Base of Xinjiang Major Diseases Research, The First Affiliated Hospital of Xinjiang Medical University Urumqi 830054, Xinjiang, P.R. China. 2. Department of Immunology, Xinjiang Medical University Urumqi 830011, Xinjiang, P.R. China. 3. Department of Hepatopancreatobiliary Surgery, Affiliated Hospital of Qinghai University 251 Xining Road, Xining 810000, Qinghai, China. 4. 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.
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%. CONCLUSION: 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.
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 infectedhuman and dog. Moreover, EgG1Y162 antigen was phylogenetically closest to EmY162 antigen, with the similarity over 90%. CONCLUSION: 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: Conan Chow; Charles G Gauci; Gulay Vural; David J Jenkins; David D Heath; Mara C Rosenzvit; Majid Fasihi Harandi; Marshall W Lightowlers Journal: Exp Parasitol Date: 2008-02-02 Impact factor: 2.011
Authors: M W Lightowlers; C G Gauci; C Chow; D R Drew; S M Gauci; D D Heath; D C Jackson; D L Dadley-Moore; A J Read Journal: Int J Parasitol Date: 2003-09-30 Impact factor: 3.981