title: [Construction and expression of a fusion gene of human heptoma peptide with human heat shock protein 70].
abstract: OBJECTIVE
  To construct and express a fusion gene of human heptoma peptide (EPVTKAEML) with human heat shock protein 70 (HSP70).
  
  
  METHODS
  A cDNA fragment encoding EPVTKAEML was added to 3 terminus of human HSP70 gene by PCR amplification. The PCR products of fusion gene were cloned into pET-28a(+)vector. The recombinant plasmid pET-28a(+)/EPVTKAEML-HSP70 was identified by enzyme digestion analysis and sequencing, and then it was transformed into E.coli BL21(DE3) through IPTG induction to express the target protein bearing His tag.
  
  
  RESULTS
  A fragment of about 2.0 kb was amplified by PCR. Sequence analysis revealed that the sequence of EPVTKAEML was connected successfully to 3 terminus of human HSP70. Enzyme digestion analysis showed the fusion gene was cloned into pET-28a(+). SDS-PAGE showed that a relative molecular mass 72 000 fusion protein was expressed.
  
  
  CONCLUSIONS
  The fusion gene of EPVTKAEML-HSP70 has been successfully constructed and expressed in E.coli BL21(DE3).
authors.affiliation: Center of Basic Medicine and Teaching Experiment, Fourth Military Medical University, Xi\rquote an 710032, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Qu
authors.forename: Jia-hai
authors.initials: JH
authors.lastname: Ma
authors.forename: Yong
authors.initials: Y
authors.lastname: Huang
authors.forename: Li-bing
authors.initials: LB
authors.lastname: Liu
authors.forename: Ru
authors.initials: R
authors.lastname: Tie
authors.forename: Fang-e
authors.initials: FE
authors.lastname: Liu
authors.forename: Xiao-jun
authors.initials: XJ
authors.lastname: Huang
mesh_terms: D006528:Carcinoma, Hepatocellular / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics / Q000378:metabolism*; D010455:Peptides / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D010455:Peptides; D011993:Recombinant Fusion Proteins