Experimental bias in the evaluation of the cellular transient expression in DNA co-transfection experiments.
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.
Bergeron
D
D
Département de Microbiologie et Immunologie, Faculté de Médecine, Université de Montréal, Québec, Canada.
Barbeau
B
B
Léger
C
C
Rassart
E
E
D000818:Animals; D016477:Artifacts; D001358:Avian Sarcoma Viruses / Q000235:genetics; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004274:DNA, Recombinant / Q000235:genetics / Q000494:pharmacology*; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics; D005069:Evaluation Studies as Topic; D015870:Gene Expression / Q000187:drug effects*; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D005814:Genes, Viral; D005822:Genetic Vectors / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012091:Repetitive Sequences, Nucleic Acid; D015203:Reproducibility of Results; D013539:Simian virus 40 / Q000235:genetics; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics; D015533:Transcriptional Activation; D014162:Transfection; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics*
D004274:DNA, Recombinant; D004268:DNA-Binding Proteins; D051789:Proto-Oncogene Protein c-fli-1; D011518:Proto-Oncogene Proteins; D011993:Recombinant Fusion Proteins; D015534:Trans-Activators; D001616:beta-Galactosidase