title: [Expression and bioassay of rubella virus E1-374 glycoprotein in yeast cells].
abstract: OBJECTIVE
  To express the rubella virus E1-374 glycoprotein in Pichia pastoris and study the immunogenecity of the recombinant protein.
  
  
  METHODS
  The cDNA of protein E1-374 was cloned into the expression vector pGAPZalphaA and transformed into Pichia pastoris GS115 cells by electrotransfection. The expressed protein was confirmed by indirect immunofluorescence and demonstrated immunoreactivity by Western Blot. Rubella virus IgG antibody was assayed with ELISA after mice were inmmunized by E1-374 glycoprotein.
  
  
  RESULTS
  SDS-PAGE analysis and Western Blot analysis of E1-374 protein revealed this protein to be 46.89 x 10(3). Antiserum (1:100) and E1-374 (5.5 microg/ml) was chosen for ELISA optimization. The intra-assay coefficient of variation for the ELISA was 0.36%-12.45%.
  
  
  CONCLUSIONS
  Protein E1-374 was highly expressed in Pichia pastoris cells, and it was a good choice to prepare rubella virus recombinant protein vaccines.
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Zhen-Mei
authors.initials: ZM
authors.lastname: Li
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Hong-Ling
authors.initials: HL
authors.lastname: Wen
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Lin
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Cheng-Xi
authors.initials: CX
authors.lastname: Sun
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Fu-Lu
authors.initials: FL
authors.lastname: Chu
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Xiao-Jing
authors.initials: XJ
authors.lastname: Yuan
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Yan-Yan
authors.initials: YY
authors.lastname: Song
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Hong-Zhi
authors.initials: HZ
authors.lastname: Xu
authors.affiliation: Department of Virology, School of Public Health, Key Laboratory for Experimental Teratology of the Ministry of Education, Shandong University, Jinan 250012, China.
authors.forename: Zhi-Yu
authors.initials: ZY
authors.lastname: Wang
mesh_terms: D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D015870:Gene Expression; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012412:Rubella virus / Q000235:genetics* / Q000276:immunology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C061826:E1 envelope protein, Rubella virus