Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

A PCR-based assay for reporter gene expression.

P Sperisen1, S M Wang, P Reichenbach

  • 1ISREC, Epalinges, Switzerland.

PCR Methods and Applications
|February 1, 1992
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

The influence of land use change on landslide susceptibility zonation: the Briga catchment test site (Messina, Italy).

Environmental management·2014
Same author

Search for new physics with a dijet plus missing E(T) signature in pp collisions at √s=1.96  TeV.

Physical review letters·2011
Same author

Electric and magnetic excitation of coherent magnetic plasmon waves in a one-dimensional meta-chain.

Optics express·2010
Same author

Measurement of the W+ W- production cross section and search for anomalous WWγ and WWZ couplings in pp collisions at square root(s)=1.96  TeV.

Physical review letters·2010
Same author

Search for WW and WZ resonances decaying to electron, missing E(T), and two jets in pp collisions at square root(s) = 1.96  TeV.

Physical review letters·2010
Same author

Search for pair production of supersymmetric top quarks in dilepton events from pp collisions at sqrt[s]=1.96  TeV.

Physical review letters·2010

This study introduces a sensitive method using polymerase chain reaction (PCR) to quantify reporter gene expression. The new assay significantly enhances the detection of cis-acting regulatory elements compared to traditional methods.

Area of Science:

  • Molecular Biology
  • Gene Regulation
  • Biotechnology

Background:

  • Transient transfection is crucial for identifying cis-acting regulatory elements by measuring reporter gene expression.
  • Current mRNA quantification methods like RNase protection and S1 mapping lack sensitivity compared to protein assays.
  • There is a need for more sensitive mRNA quantification techniques in gene expression studies.

Purpose of the Study:

  • To develop a highly sensitive method for quantifying reporter gene expression using the polymerase chain reaction (PCR).
  • To improve the detection of cis-acting regulatory elements through enhanced gene expression analysis.
  • To establish a reliable and sensitive assay for measuring mRNA levels.

Main Methods:

  • Co-transfection of cells with a test construct and a reference plasmid containing a modified beta-globin gene.

Related Experiment Videos

  • Utilizing the polymerase chain reaction (PCR) to amplify cDNA from both the test and reference constructs.
  • Quantifying gene expression by measuring the ratio of amplified cDNA signals.

    • Main Results:

    • Developed a novel PCR-based system for quantifying rabbit beta-globin reporter gene expression.
    • Demonstrated that the ratio of amplified cDNA signals provides a highly reliable measure of gene expression.
    • Achieved a sensitivity at least 1000-fold higher than RNase protection assays.

    Conclusions:

    • The developed PCR-based assay offers a significant advancement in sensitivity for quantifying reporter gene expression.
    • This method provides a reliable and highly sensitive tool for identifying cis-acting regulatory elements.
    • The enhanced sensitivity facilitates more accurate and robust gene expression analysis in molecular biology research.