title: [Cloning, expression of TSO45W-4B gene from Taenia solium and preparation of its polyclonal antibody].
abstract: OBJECTIVE
  To clone and express TSO45W-4B gene of Taenia solium, and prepare the rabbit antiserum against the recombinant protein TSO45W-4B.
  
  
  METHODS
  The gene encoding TSO45W-4B antigen was synthesized and cloned into the expression vector pGEX-1lambdaT. The recombinant plasmid was transformed into Escherichia coli ArcticExpress (DE3), and followed by the expression of the protein induced by IPTG. The recombinant protein was purified by GST affinity chromatography and analyzed by SDS-PAGE. The rabbits were immunized with the purified recombinant protein. The titer of the antibody against TSO45W-4B was detected by ELISA. The specificity of anti-TSO45W-4B antibody was determined by Western blotting.
  
  
  RESULTS
  It was demonstrated that TSO45W-4B gene (351 bp) was synthesized. The gene was inserted into pGEX-1lambdaT and confirmed by restriction enzyme digestion and DNA sequencing. SDS-PAGE analysis showed that the relative molecular mass (M(r)) of the expressed recombinant protein was approximately 40 000. The purity of the recombinant protein with an affinity chromatography column was about 90%. The titer of the antibody against TSO45W-4B was 1: 512 000. The recombinant protein reacted with the anti-TSO45W-4B antibody by Western blotting.
  
  
  CONCLUSIONS
  The TSO45W-4B gene of Taenia solium is efficiently expressed in E. coli. The recombinant protein TSO45W-4B and its rabbit antiserum are successfully prepared.
authors.forename: Bi-Ying
authors.initials: BY
authors.lastname: Zhou
authors.forename: Ling
authors.initials: L
authors.lastname: Zhou
authors.forename: Mei-Chen
authors.initials: MC
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Liu
authors.forename: Xi
authors.initials: X
authors.lastname: Zhang
mesh_terms: D000818:Animals; D000909:Antibodies, Helminth; D000947:Antigens, Helminth / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D017238:Genes, Helminth; D005822:Genetic Vectors; D007106:Immune Sera; D008297:Male; D011817:Rabbits; D041201:Taenia solium / Q000235:genetics* / Q000276:immunology*
chemical_list: D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D007106:Immune Sera