title: Reporter gene expression at single-cell level characterized with real-time RT-PCR, chemiluminescence, fluorescence, and electrochemical imaging.
abstract: mRNA from single cells was quantified using real-time RT-PCR after recording the address and reporter protein activity with chemiluminescence, fluorescence, and electrochemical techniques, using luciferase, green fluorescent protein, and secreted alkaline phosphatase. mRNA copy number ranging from below 10(3) to 10(7) in single cells showed a lognormal distribution for both externally introduced reporter genes and internally expressed genes. The fluctuation in the gene expression decreased with the increase of the number of cells picked but did not decrease with the increase of mRNA copy number per cell. We found that the correlation coefficients for mRNA and protein expression in logarithmic plot at single-cell level were much lower than 1.00.
authors.affiliation: Graduate School of Environmental Studies, Tohoku University, Aramaki-Aoba, Sendai, Japan. shiku@bioinfo.che.tohoku.ac.jp
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Shiku
authors.forename: Daisuke
authors.initials: D
authors.lastname: Okazaki
authors.forename: Junya
authors.initials: J
authors.lastname: Suzuki
authors.forename: Yasufumi
authors.initials: Y
authors.lastname: Takahashi
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Murata
authors.forename: Hidetaka
authors.initials: H
authors.lastname: Akita
authors.forename: Hideyoshi
authors.initials: H
authors.lastname: Harashima
authors.forename: Kosuke
authors.initials: K
authors.lastname: Ino
authors.forename: Tomokazu
authors.initials: T
authors.lastname: Matsue
mesh_terms: D000469:Alkaline Phosphatase / Q000032:analysis / Q000235:genetics; D002477:Cells / Q000378:metabolism*; D005453:Fluorescence; D015870:Gene Expression; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D049449:Luminescence; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D008156:Luciferases; D000469:Alkaline Phosphatase