title: [Construction of middle fragment deletion mutant with improved gene splicing by overlap extension].
abstract: OBJECTIVE
  To construct a phosphatidylinositol 4-kinase beta (PI4K-beta) mutant with the 325th to 373rd amino acid codons deleted, and try to develop a simple method for constructing middle fragment deletion mutant.
  
  
  METHODS
  In line with the mechanism of gene splicing by overlap extension(SOE), an additional PCR was used to get the PI4K-beta mutant in which the 325th to 373rd amino acid codons were deleted. Then the mutated gene was cloned into pCMV-Tag4A mammalian expression vector.
  
  
  RESULTS
  A mutant with the 325th to 373rd amino acid codons deleted was constructed successfully.
  
  
  CONCLUSIONS
  The improved SOE is a very effective and reliable method to construct middle fragment deletion mutant. It is worthy to be popularized.
authors.affiliation: National Laboratory of Medical Genetics, Central South University, Changsha, Hunan, PR China.
authors.forename: Chen-hua
authors.initials: CH
authors.lastname: Li
authors.forename: Hai-yan
authors.initials: HY
authors.lastname: Fang
authors.forename: Xiao-yun
authors.initials: XY
authors.lastname: Deng
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.forename: Duo
authors.initials: D
authors.lastname: Zheng
authors.forename: Jia-hui
authors.initials: JH
authors.lastname: Xia
mesh_terms: D019870:1-Phosphatidylinositol 4-Kinase / Q000235:genetics*; D001483:Base Sequence; D005822:Genetic Vectors / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D015202:Protein Engineering / Q000379:methods; D011994:Recombinant Proteins / Q000235:genetics; D017384:Sequence Deletion
chemical_list: D011994:Recombinant Proteins; D019870:1-Phosphatidylinositol 4-Kinase