A PCR based assay for reporter gene expression

Peter Sperisen, San Ming Wang, Patrick Reichenbach, Markus Nabholz

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Transient transfection is a widely used tool for the identification of cis-acting regulatory elements. These elements are detected by their effect on the expression of a reporter gene, which is quantified by measuring the reporter gene product in the form of mRNA, protein (hGH), or enzymes (CAT, Iuciferase). Measurements of mRNA levels have several advantages over enzyme or protein assays. However, mRNA quantification by RNase protection or S1 mapping has considerably lower signal-to-background ratio than protein assays and is therefore less sensitive. In this paper we report the development of a system that takes advantage of the polymerase chain reaction (PCR) to quantify rabbit β-globin reporter gene expression. Cells are co-transfected with constructs whose activity is to be tested and a reference plasmid with a small deletion in the second exon of the β-globin gene. We show that the ratio of the two amplified cDNA signals is a highly reliable measure of test gene expression. The sensitivity of this assay is at least 1000-fold higher than RNase protection.

Original languageEnglish (US)
Pages (from-to)164-170
Number of pages7
JournalGenome Research
Volume1
Issue number3
DOIs
StatePublished - Jan 1 1992

Fingerprint

Reporter Genes
Globins
Ribonucleases
Gene Expression
Polymerase Chain Reaction
Messenger RNA
Proteins
Chloramphenicol O-Acetyltransferase
Transfection
Exons
Plasmids
Complementary DNA
Rabbits
Enzymes
Genes

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Sperisen, P., Wang, S. M., Reichenbach, P., & Nabholz, M. (1992). A PCR based assay for reporter gene expression. Genome Research, 1(3), 164-170. https://doi.org/10.1101/gr.1.3.164

A PCR based assay for reporter gene expression. / Sperisen, Peter; Wang, San Ming; Reichenbach, Patrick; Nabholz, Markus.

In: Genome Research, Vol. 1, No. 3, 01.01.1992, p. 164-170.

Research output: Contribution to journalArticle

Sperisen, P, Wang, SM, Reichenbach, P & Nabholz, M 1992, 'A PCR based assay for reporter gene expression', Genome Research, vol. 1, no. 3, pp. 164-170. https://doi.org/10.1101/gr.1.3.164
Sperisen P, Wang SM, Reichenbach P, Nabholz M. A PCR based assay for reporter gene expression. Genome Research. 1992 Jan 1;1(3):164-170. https://doi.org/10.1101/gr.1.3.164
Sperisen, Peter ; Wang, San Ming ; Reichenbach, Patrick ; Nabholz, Markus. / A PCR based assay for reporter gene expression. In: Genome Research. 1992 ; Vol. 1, No. 3. pp. 164-170.
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