title: [Molecular cloning of recombinant fibronectin EDA and EDB fusion protein].
abstract: OBJECTIVE
  Construct a recombinant plasmid pET28a-EDA-EDB, prepare the fusion EDA-EDB protein.
  
  
  METHODS
  For the production of recombinant fibronectin EDA-EDB in Escherichia coli, the EDA and EDB segments were separated from pGEM2-EDA/EDB and recomposed with two additional amino acids, then cloned into the expression vector pET28a. pET system to express EDA-EDB fusion protein and 6 x His/Ni-NTA system to purify it in a single step were used. Western blotting confirmed the purified protein.
  
  
  RESULTS
  The EDA and EDB segments were ligated and inserted into pET28a vector. EDA-EDB fusion protein was highly expressed in Escherichia coli BL21 (DE3). Afterwards, it was purified by Ni-NTA resin and verified by western blotting.
  
  
  CONCLUSIONS
  EDA-EDB fusion protein can be expressed in pET system and purified by 6 x His/Ni-NTA system.
authors.affiliation: Department of Forensic Medicine, Shanghai Medical School, Fudan University, Shanghai 200032. aimin-xue@yahoo.com.cn
authors.forename: Ai-min
authors.initials: AM
authors.lastname: Xue
authors.forename: Hua
authors.initials: H
authors.lastname: Wang
authors.forename: Rong
authors.initials: R
authors.lastname: Ye
mesh_terms: D015153:Blotting, Western; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D005353:Fibronectins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins