Experimental bias in the evaluation of the cellular transient expression in DNA co-transfection experiments

Cell Mol Biol Res. 1995;41(3):155-9.

Abstract

beta-Galactosidase (beta-gal) expressing vectors are commonly used to standardize the transfection efficiency in transient expression experiments. In the Chinese hamster ovary (CHO) cell line, we transfected beta-gal expressing vectors in combination with different plasmid DNAs. We reported here that the presence of specific DNAs led to statistically significant variations in the beta-gal expression level. Therefore, the measure of beta-gal activity is not necessarily an accurate method to monitor transfection efficiency, and its use to normalize the expression from reporter genes could be questionable.

MeSH terms

  • Animals
  • Artifacts*
  • Avian Sarcoma Viruses / genetics
  • CHO Cells / metabolism
  • Cricetinae
  • Cricetulus
  • DNA, Recombinant / genetics
  • DNA, Recombinant / pharmacology*
  • DNA-Binding Proteins / biosynthesis
  • DNA-Binding Proteins / genetics
  • Evaluation Studies as Topic
  • Gene Expression / drug effects*
  • Gene Expression Regulation
  • Genes, Reporter*
  • Genes, Viral
  • Genetic Vectors / genetics*
  • Promoter Regions, Genetic
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins*
  • Recombinant Fusion Proteins / biosynthesis*
  • Recombinant Fusion Proteins / genetics
  • Repetitive Sequences, Nucleic Acid
  • Reproducibility of Results
  • Simian virus 40 / genetics
  • Trans-Activators / biosynthesis
  • Trans-Activators / genetics
  • Transcriptional Activation*
  • Transfection*
  • beta-Galactosidase / biosynthesis
  • beta-Galactosidase / genetics*

Substances

  • DNA, Recombinant
  • DNA-Binding Proteins
  • Proto-Oncogene Protein c-fli-1
  • Proto-Oncogene Proteins
  • Recombinant Fusion Proteins
  • Trans-Activators
  • beta-Galactosidase