title: [Construction of Escherichia coli gene knock-out mutants for engineering of fatty acid metabolism].
abstract: OBJECTIVE
  Gene knock out technique is very important for gene function study. We developed a simple and efficient method to knock out chromosomal genes in Escherichia coli.
  
  
  METHODS
  Using the Escherichia coli Keio single-mutant library, we combined Red homology recombination with P1 phage transduction and developed a method to knock out the genes in Escherichia coli MG1655.
  
  
  RESULTS
  We obtained beta-oxidation mutants deltafadD, deltafadE and deltafadD-deltafadE and fatty acid synthesis mutants deltafabH, deltafabF and deltafabH-deltafabF. There were no obvious growth changes between deltafadD or deltafadE mutant strains and MG1655. However, deltafabH and deltafabH-deltafabF mutant strains grew much slower than the wild type strain. The fatty acid contents in deltafadD, deltafadE and deltafadD-deltafadE were 18.2 mg/L, 20.0 mg/L and 19.2 mg/L respectively, higher than 17.5 mg/L in wild type. The fatty acid contents in deltafabH, deltafabF and deltafabH-deltafabF were 12.6 mg/L, 15.2 mg/L and 11.2mg/L respectively, lower than that in wild type.
  
  
  CONCLUSIONS
  Using Keio mutant library, P1 phage transduction and resistant gene elimination, we have established a simple and efficient method for gene knock-out in Escherichia coli.
authors.affiliation: Institute of Microbiology Chinese Academy of Sciences, Beijing 100190, China. qujingqiu1986@163.com
authors.forename: Jingqiu
authors.initials: J
authors.lastname: Qu
authors.forename: Cuihua
authors.initials: C
authors.lastname: Liu
authors.forename: Weifeng
authors.initials: W
authors.lastname: Liu
authors.forename: Yong
authors.initials: Y
authors.lastname: Tao
authors.forename: Kun
authors.initials: K
authors.lastname: Zhu
mesh_terms: D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism*; D055786:Gene Knockout Techniques / Q000379:methods*; D009154:Mutation
chemical_list: D029968:Escherichia coli Proteins; D005227:Fatty Acids