title: [Construction and identification of the recombinant prokaryotic expression plasmid containingbetapep-25 peptide].
abstract: OBJECTIVE
  To synthesize antiangiogenic peptide fragment of betapep-25, to construct and identify the recombinant prokaryotic expression plasmid containing betapep-25 peptide.
  
  
  METHODS
  The fragment encoding betapep-25 peptide was designed and synthesized artificially and was cloned into vector pGEM-T easy after being identified by sequencing. After being digested by Nae I and BamH I, T-betapep-25 peptide fragment was cloned into recombinant vector pBV220-NT4, which was digested by Nae I and BamH I. The constructed recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 was digested using BamH I, EcoR I and BamH I, respectively.
  
  
  RESULTS
  The sequcence of betapep-25 synthesized artificially and analyzed by digestion was consistent with the published results. Fragments of 4,030 bp, 364 bp and 3,666 bp were obtained after digestion of recombinant prokaryotic expression plasmid pBV220-NT4-betapep-25 using BamH I, EcoR I and BamH I, respectively, which demonstrated the successful cloning and construction of recombinant prokaryotic expression plasmid.
  
  
  CONCLUSIONS
  The successful construction recombinant prokaryotic expression plasmid expressing betapep-25 provides a foundation for further research on its mechanism of anti-tumor activity in vivo and in vitro.
authors.affiliation: Department of Medical Oncology of the First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, China. lienxiao@csco.org.cn
authors.forename: En-xiao
authors.initials: EX
authors.lastname: Li
authors.forename: Shan-xi
authors.initials: SX
authors.lastname: Liu
authors.forename: Guang-xiao
authors.initials: GX
authors.lastname: Yang
authors.forename: Quan-ying
authors.initials: QY
authors.lastname: Wang
authors.forename: Yin-ying
authors.initials: YY
authors.lastname: Wu
authors.forename: Fan
authors.initials: F
authors.lastname: Shi
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D010455:Peptides / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D011387:Prokaryotic Cells / Q000378:metabolism*; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D020798:Two-Hybrid System Techniques
chemical_list: D010455:Peptides; D011506:Proteins; D011994:Recombinant Proteins; C111993:betapep-25 protein, synthetic