title: [Optimizing expression and antibody preparation of recombinant Streptococcus mutans surface protein].
abstract: OBJECTIVE
  The soluble protein recombinant Streptococcus mutans surface protein (rPAc) was expressed in Escherichia coli (E.coli) after the optimization of inducing conditions. The antiserum against rPAc was obtained by immunizing mice with the purified rPAc.
  
  
  METHODS
  The soluble expression of rPAc in E. coli was further optimized by means of different culture conditions. Polyclonal antibody was made by immunizing mice with purified rPAc. Western blot and enzyme linked immunosorbent assay (ELISA) were carried out to identify the immunocompetence of the antibody.
  
  
  RESULTS
  The highest soluble expression level of rPAc was obtained at Luria-Bertani (LB) medium (pH=7.2) when optical density (OD600nm) was 0.6 after being induced at 30 degrees C for 4 h and the concentration of isopropyl beta-D-1-thigalactopyranoside (IPTG) was 1.0mmol x L(-1). The titer of the mice antiserum against rPAc was about 1:6000 by ELISA analysis, and rPAc could be specifically recognized by Western blot analysis.
  
  
  CONCLUSIONS
  This study proved that rPAc can be effectively expressed as a soluble form in E. coli, and the high specific polyclonal antibody of rPAc was proved to be prepared, which shed light on further research of DNA prime-protein boost inoculation.
authors: Dept. of Operative Dentistry and Endodontics, Stomatological Hospital of Hangzhou, Hangzhou 310006, China.
authors: Jie
authors: J
authors: Jin
authors: Mingwen
authors: M
authors: Fan
authors: Yuhong
authors: Y
authors: Li
mesh_terms: D000818:Animals; D000906:Antibodies; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli; D008565:Membrane Proteins; D051379:Mice; D011994:Recombinant Proteins; D013295:Streptococcus mutans
chemical_list: D000906:Antibodies; D008565:Membrane Proteins; D011994:Recombinant Proteins