title: [The expression, purification and characterization of β-hCG protein in E.coli].
abstract: OBJECTIVE
  To obtain β-chain human chorionic gonadotropin (β-hCG) fusion protein (β-hCG/GST) and identify its antigenicity.
  
  
  METHODS
  The full-length gene of β-hCG was amplified by PCR. The PCR product was cloned into pET-42a prokaryotic expression vector to construct the recombinant plasmid pET-42a-β-hCG, and then it was transformed into BL21 (DE3) for β-hCG/GST fusion protein expression under IPTG induction. After SDS-PAGE assay, the fusion protein was purified by affinity chromatography and identified by Western blotting. The antigenicity of the purified fusion protein was characterized by ELISA.
  
  
  RESULTS
  The β-hCG gene we obtained had an identical sequence to that retrieved in GenBank. The prokaryotic expression vector pET-42a-β-hCG was successfully constructed as confirmed by enzyme digestion and DNA sequencing. Both Western blotting and ELISA demonstrated that the purified β-hCG fusion protein had satisfactory antigenicity.
  
  
  CONCLUSIONS
  The purified β-hCG/GST fusion protein with satisfactory antigenicity has been obtained, which will facilitate further study on active anti-tumor immunotherapy targeting β-hCG.
authors: Department of Urology, General Hospital of PLA, Beijing 100853, China. wayewong261@yahoo.cn
authors: Wei
authors: W
authors: Wang
authors: Xiaotao
authors: X
authors: Yin
authors: Yunqi
authors: Y
authors: Li
authors: Renli
authors: R
authors: Tian
authors: Hao
authors: H
authors: Wu
authors: Jinqi
authors: J
authors: Yan
authors: Jiangping
authors: J
authors: Gao
authors: Jiyun
authors: J
authors: Yu
mesh_terms: D018997:Chorionic Gonadotropin, beta Subunit, Human / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins
chemical_list: D018997:Chorionic Gonadotropin, beta Subunit, Human; D011993:Recombinant Fusion Proteins