title: [Development of a mouse cell line containing stably integrated copies of pMCLacI/Neo plasmid: a model for studying mutations in vitro].
abstract: OBJECTIVE
  To establish a suitable model for studying the different mechanisms of mutation between expressed and non-expressed genes in mammalian cells.
  
  
  METHODS
  The NIH3T3 cells were transfected with the linearized pMCLacI/Neo DNAs by liposome-mediated transfection, and grew in the presence of G418. One drug resistant cell clone was selected to proliferate and to be analyzed with Southern blot and RT-PCR analyses on its genomic DNAs.
  
  
  RESULTS
  (1) Multiple copies of pMCLacI/Neo plasmid DNA were intactly integrated in the genomic DNAs of the cell clone. (2) One of lac I target genes in the integrated plasmid could be transcribed in the NIH3T3 cells while the other could not. (3) The pMCLacI/Neo plasmid DNA could be efficiently rescued from the genomic DNAs of the cell clone with the average rescue efficiency of 410 cfu/microg DNA.
  
  
  CONCLUSIONS
  The NIH3T3 cell line containing copies of a stably integrated pMCLacI/Neo has been established. The two lacI target genes in the cell line could imitate the functional states of expressed and non-expressed genes in mammalian cells respectively. The cell line will be a useful model for studying the different mechanisms of mutation between expressed and non-expressed genes in mammalian cells.
authors.affiliation: Department of Medical Genetics, Second Military Medical University, Shanghai, 200433 P.R. China. Huaixing@bigfoot.com.
authors.forename: Y
authors.initials: Y
authors.lastname: Lu
authors.forename: H
authors.initials: H
authors.lastname: Li
authors.forename: J
authors.initials: J
authors.lastname: Fu
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D051379:Mice; D009154:Mutation; D010957:Plasmids; D012097:Repressor Proteins / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D056968:Lac Repressors; D012097:Repressor Proteins