title: [Clonning and expression of procalcitonin and preparation of its polyclonal antibody].
abstract: OBJECTIVE
  To clone, express the procalcitonin (PCT) and prepare its polyclonal antibody.
  
  
  METHODS
  We designed 10 DNA primers according to the encoding sequence of human PCT gene from GenBank, amplified PCT gene by PCR and constructed the recombinant vector pGEX-KG-PCT. Then the fusion protein, PCT-GST, was expressed in E.coli and purified by affinity chromatography. The corresponding polyclonal antibody was produced in the New Zealand white rabbits immunized with the fusion protein, and its biological activity was detected by Western blotting. The specificity and titer of the polyclonal antibody was identified by agar gel immunodiffusion test.
  
  
  RESULTS
  We successfully obtained the full-length PCT gene and expressed the fusion PCT-GST in E.coli. SDS-PAGE showed that relative molecular mass (Mr;) was about 36 000. Agar gel immunodiffusion test revealed that the prepared polyclonal antibody could specially recognized PCT and its titer was 1:4.
  
  
  CONCLUSIONS
  The PCT is successfully cloned, expressed and purified. The high specific polyclonal antibody of PCT is prepared.
authors: School of Medicine, Wuhan University, China. 53820789@qq.com
authors: Qian
authors: Q
authors: Tan
authors: Weiqun
authors: W
authors: Tian
authors: Li
authors: L
authors: Wang
authors: Wei
authors: W
authors: Peng
authors: Fan
authors: F
authors: Chen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology; D002116:Calcitonin / Q000235:genetics* / Q000276:immunology*; D015740:Calcitonin Gene-Related Peptide; D003001:Cloning, Molecular; D015870:Gene Expression; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011498:Protein Precursors / Q000235:genetics* / Q000276:immunology*; D011817:Rabbits; D011993:Recombinant Fusion Proteins
chemical_list: D000906:Antibodies; C000606589:CALCA protein, human; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D002116:Calcitonin; D015740:Calcitonin Gene-Related Peptide