title: [Cloning and expression of PbTIP analogous gene from Plasmodium berghei ANKA and preparation of its polyclonal antibody].
abstract: OBJECTIVE
  To clone and express a novel protein analogous to TIP (T cell immunomodulatory protein) of Plasmodium berghei ANKA and prepare its polyclonal antibody.
  
  
  METHODS
  The PbTIP encoding nucleotide sequence was searched from the Plasmodium berghei genomic database and amplified by PCR. The gene was sub-cloned into prokaryotic expression vector pGEX4T-1 and expressed in E.coli BL21 (DE3). After induction by IPTG, the expression of PbTIP-GST fusion protein was characterized by SDS-PAGE and Western blotting. The inclusion bodies of GST-PbTIP fusion protein were injected into BALB/c mouse. Anti-sera were identified by indirect fluorescent antibody test and western blotting.
  
  
  RESULTS
  The PbTIP-GST fusion protein was successfully expressed in the form of inclusion bodies, by controlling the temperature and concentration of IPTG. Anti-PbTIP-GST sera were acquired with high titer. The sera specifically recognized the PbTIP with a band of 60 000 in P.berghei infected erythrocyte protein.
  
  
  CONCLUSIONS
  PbTIP/GST fusion protein and polyclonal antibody have been obtained.
authors.forename: Ying-Hui
authors.initials: YH
authors.lastname: Li
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhao
authors.forename: Zhong-Xiang
authors.initials: ZX
authors.lastname: Liu
authors.forename: Yu-Xiao
authors.initials: YX
authors.lastname: Huang
authors.forename: Zhang-Xiang
authors.initials: ZX
authors.lastname: Mao
authors.forename: Cai-Fang
authors.initials: CF
authors.lastname: Xue
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D003001:Cloning, Molecular; D015870:Gene Expression; D051379:Mice; D008807:Mice, Inbred BALB C; D010962:Plasmodium berghei / Q000235:genetics* / Q000378:metabolism; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D011994:Recombinant Proteins / Q000276:immunology; D037182:T-Cell Antigen Receptor Specificity
chemical_list: D000913:Antibodies, Protozoan; D015800:Protozoan Proteins; D011994:Recombinant Proteins