title: [Expression and purification of HPV16 E2 protein and preparation of its antiserum].
abstract: OBJECTIVE
  To express and purify the human papillomavirus type 16 (HPV16) E2 protein in prokaryotic bacteria and prepare the antiserum of HPV16 E2.
  
  
  METHODS
  After amplified by PCR, HPV16 E2 was inserted into pET21b vector. The recombinant pET21b-HPV16E2 vector was transfected into E.coli BL21 (DE3). Expression product was identified after induction. Through purification, denaturation and renaturation, soluble protein was obtained. With the HPV16 E2 protein, we immunized BALB/c mice and examined mouse IFN-γ, CD4(+); T cells, CD8(+); T cells, CD4/CD8 ratio and antiserum titer.
  
  
  RESULTS
  Restriction digestion and DNA sequencing showed pET21b-HPV16E2 was constructed successfully. Relative molecular mass (Mr;) of HPV16 E2 was 42 000 in SDS-PAGE and the specificity of the protein was confirmed with Western blotting. The antiserum could specifically bind with HPV16 E2 protein. In the immunized BALB/c mice, antiserum titre, CD4(+); T cell count and CD4/CD8 ratio increased, while mouse IFN-γ did not change obviously.
  
  
  CONCLUSIONS
  Soluble HPV16 E2 protein was obtained successfully. The antiserum of high titer against HPV16 E2 was prepared in mice.
authors.affiliation: Department of Microbiology, Harbin Medical University, Harbin, China.
authors.forename: Yuhui
authors.initials: Y
authors.lastname: Sun
authors.forename: Yanjun
authors.initials: Y
authors.lastname: Zhang
authors.forename: Mingming
authors.initials: M
authors.lastname: Liu
authors.forename: Liping
authors.initials: L
authors.lastname: Tang
authors.forename: Guanghong
authors.initials: G
authors.lastname: Zhang
authors.forename: Lanlan
authors.initials: L
authors.lastname: Wei
authors.forename: Hongxi
authors.initials: H
authors.lastname: Gu
authors.forename: Qinglong
authors.initials: Q
authors.lastname: Shang
mesh_terms: D000818:Animals; D016516:CD4-CD8 Ratio; D016207:Cytokines; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D007106:Immune Sera / Q000096:biosynthesis*; D051379:Mice; D008807:Mice, Inbred BALB C; D009856:Oncogene Proteins, Viral / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology; D017422:Sequence Analysis, DNA; D014452:Ubiquitins
chemical_list: D016207:Cytokines; D004268:DNA-Binding Proteins; C054855:E2 protein, Human papillomavirus type 16; C489550:G1p2 protein, mouse; D007106:Immune Sera; D009856:Oncogene Proteins, Viral; D011994:Recombinant Proteins; D014452:Ubiquitins