title: [Cloning, expression and identification of Plasmodium vivax Duffy binding protein region II of central China isolate].
abstract: OBJECTIVE
  To clone a Plasmodium vivax Duffy binding protein critical functional region II (PvDBPII) gene of central China isolate, and to express and identify the recombinant PvDBPII protein in vitro.
  
  
  METHODS
  PCR was performed to amplify PvDBPII from P. vivax DNA of a central China isolate and the PCR product was inserted into pET28a(+) vector. pET28a-PvDBPII recombinant plasmid was constructed and transformed into E. coli host BL21 (DE3+). IPTG was used to induce the recombinant PvDBPII protein fused with His tag, and the protein was purified by His-NTA affinity chromatography. The recombinant protein was identified by SDS-PAGE and Western blot.
  
  
  RESULTS
  The PCR product of PvDBPII gene was about 1,1 kb, meeting the expectation of predicted fragment size. The recombinant pET28a-PvDBPII plasmid was verified by sequencing that the insertion was correct both in direction and in frame, but with 4 non-synonymous mutations compared to reference P. vivax strain Sal-I. SDS-PAGE, and Western blot analysis showed that the recombinant PvDBPII protein was about 44 kDa, and could be recognized by pooled sera from vivax malaria patients.
  
  
  CONCLUSIONS
  The PvDBPII gene of central China isolate is successfully cloned, and recombinant PvDBPII is expressed, thereby providing opportunity for further study on PvDBPII.
authors.affiliation: Department of Microbiology and Parasitology, Bengbu Medical College, Bengbu 233030, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Liu
authors.forename: Hui
authors.initials: H
authors.lastname: Xia
authors.forename: Zhi-Yong
authors.initials: ZY
authors.lastname: Tao
authors.forename: Yong
authors.initials: Y
authors.lastname: Chen
authors.forename: Qiang
authors.initials: Q
authors.lastname: Fang
authors.forename: Xue-Mei
authors.initials: XM
authors.lastname: Wang
authors.forename: Xin
authors.initials: X
authors.lastname: Sun
authors.forename: Ying
authors.initials: Y
authors.lastname: Gao
authors.forename: Yue-Qin
authors.initials: YQ
authors.lastname: Mai
mesh_terms: D000953:Antigens, Protozoan / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001665:Binding Sites; D015153:Blotting, Western; D002681:China; D003001:Cloning, Molecular; D006801:Humans; D017780:Malaria Vaccines / Q000276:immunology; D010966:Plasmodium vivax / Q000276:immunology*; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000276:immunology
chemical_list: D000953:Antigens, Protozoan; C066001:Duffy antigen binding protein, Plasmodium; D017780:Malaria Vaccines; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins