title: [Prokaryotic expression, purification and antigenicity identification of mouse prostate stem cell antigen].
abstract: OBJECTIVE
  To amplify mouse prostate stem cell antigen (mPSCA) gene and construct a recombinant plasmid to obtain mPSCA protein and identify its antigenicity.
  
  
  METHODS
  The gene of mPSCA was amplified by RT-PCR from mouse prostate cancer cell line RM-1 with the signal peptide sequence removed. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-mPSCA, which was transformed into BL21 (DE3) for mPSCA expression. The fusion protein was purified and identified by SDS-PAGE and Western blotting. The antigenicity of the purified protein was characterized by ELISA.
  
  
  RESULTS
  The mPSCA gene was obtained with an identical sequence to that retrieved in GenBank. The prokaryotic expression vector for mPSCA was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated the antigenicity of the purified mPSCA protein.
  
  
  CONCLUSIONS
  The purified mPSCA obtained possesses good antigenicity, which will facilitate further study of immunotherapy for prostate cancer targeting PSCA.
authors.affiliation: Department of Urology, General Hospital of PLA, Beijing 100853, China. jk_dong@163.com
authors.forename: Jinkai
authors.initials: J
authors.lastname: Dong
authors.forename: Jin
authors.initials: J
authors.lastname: Luo
authors.forename: Jinqi
authors.initials: J
authors.lastname: Yan
authors.forename: Liang
authors.initials: L
authors.lastname: Zhang
authors.forename: Jiangping
authors.initials: J
authors.lastname: Gao
authors.forename: Jiyun
authors.initials: J
authors.lastname: Yu
mesh_terms: D000818:Animals; D000951:Antigens, Neoplasm / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D058851:GPI-Linked Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D005822:Genetic Vectors; D008297:Male; D051379:Mice; D009363:Neoplasm Proteins / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification; D010957:Plasmids; D011467:Prostate / Q000166:cytology
chemical_list: D000951:Antigens, Neoplasm; D058851:GPI-Linked Proteins; D009363:Neoplasm Proteins; C504399:Psca protein, mouse