title: [Prokaryotic expression, purification and biological activity analysis of recombinant β-Lactamase protein].
abstract: OBJECTIVE
  To prepare RGD4CβL fusion protein using prokaryotic expression system and evaluate the biological activity of the RGD4CβL.
  
  
  METHODS
  RGD4CβL gene was cloned into pColdII to contruct β-Lactamase prokaryotic expression vector. After transformation, the recombinant vector was induced to express recombinant protein RGD4CβL by IPTG in E.coli BL(DE3). The recombinant protein was purified by Ni-NTA resin under denaturing condition and then dialyzed to renature. The tumor cell targeting ability of the recombinant protein was analyzed by flow cytometric analysis.
  
  
  RESULTS
  After cleavage and purification, β-Lactamase moiety showed the expected size of 42 000 on Tricine-SDS-PAGE, and was further confirmed by Western blotting. Based on flow cytometric analysis, the purified protein specially targeted breast cancer cell line MCF-7.
  
  
  CONCLUSIONS
  This research successfully estiblished a method for prokaryotic expression and purification of β-lactamase. These results suggest the potential use of the protein as an agent for ADEPT.
authors.affiliation: Institute of Radiation Medicine, Chinese Academy of Medical Sciences, Peking Union Medical College, Tianjin 300192, China.
authors.forename: Xiao-liang
authors.initials: XL
authors.lastname: Zhou
authors.forename: Pei-ji
authors.initials: PJ
authors.lastname: Shi
authors.forename: Hao
authors.initials: H
authors.lastname: Wang
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000188:drug therapy; D045744:Cell Line, Tumor; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D015870:Gene Expression; D005822:Genetic Vectors; D010957:Plasmids / Q000235:genetics; D011387:Prokaryotic Cells / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*; D017422:Sequence Analysis, DNA / Q000379:methods; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D011993:Recombinant Fusion Proteins; D001618:beta-Lactamases