title: [Engineering of Escherichia coli for convenient expression of [FeFe]-hydrogenase].
abstract: OBJECTIVE
  A new method used to heterologously express [FeFe]-hydrogenase in Escherichia coli was investigated in our present study.
  
  
  METHODS
  By homologous recombination, three assistant genes (hydE, hydF and hydG) for hydrogenase were integrated into the chromosome of E. coli BW 25113-10, in which all hydrogenase genes were inactivated. A hydrogenase structural gene hydA from Clostridium butyricum was used to test the hydrogenase maturation ability of the recombined E. coli. BW 25113-13.
  
  
  RESULTS
  The corrected integration of the three assistant genes was confirmed by PCR, and RT-PCR results indicated that the three accessory genes were transcripted in the recombinant. The active expression of hydA indicated that the constitutively expressed accessory proteins could assist the maturation of the [FeFe]-hydrogenases.
  
  
  CONCLUSIONS
  A simplified [FeFe]-hydrogenase expression recombinant E. coli BW25113-13 was constructed. It would lay foundations for the functional screening of [FeFe]-hydrogenases and the construction of novel hydrogen producing pathways in E. coli.
authors.affiliation: Shanghai Key Laboratory of Agricultural Genetics and Breeding, Institute of Animal Science and Veterinary Medicine, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China. yursong@163.com
authors.forename: Ruisong
authors.initials: R
authors.lastname: Yu
authors.forename: Wenming
authors.initials: W
authors.lastname: Zong
authors.forename: Zhihua
authors.initials: Z
authors.lastname: Zhou
mesh_terms: D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D059765:Homologous Recombination; D006864:Hydrogenase / Q000096:biosynthesis* / Q000235:genetics*; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis* / Q000235:genetics*
chemical_list: D007506:Iron-Sulfur Proteins; C118830:iron hydrogenase; D006864:Hydrogenase