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Real-Time Reverse Transcription PCR as a Tool to Study Virulence Gene Regulation in Bacterial Pathogens

  • Gili Aviv
  • Ohad Gal-MorEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1734)

Abstract

Quantitative real-time PCR (qRT-PCR) is a highly sensitive and reliable method for detection and quantification of DNA. When combined with a prior stage of RNA reverse transcription to generate complementary DNA (cDNA), this is a powerful approach to determine and analyze gene transcriptional expression. Real-time quantitative reverse transcription PCR has become the gold standard method in studying genes expression and virulence regulation under various genetic backgrounds (e.g., in the absence of regulators) or environmental conditions. Here we demonstrate the utilization of this approach to study the transcriptional regulation of the conjugation pilus of the Salmonella enterica serovar Infantis virulence plasmid (pESI).

Key words

Real-time PCR Reverse transcription cDNA SYBR green dye ROX Transcription Regulation Gene expression 

Notes

Acknowledgments

The research in Gal-Mor lab is supported by a grant number 1096.39.11/2010 from the German-Israeli Foundation for Scientific Research and Development (GIF); by a grant number 999/14 from the Israel Science Foundation (ISF) and by grant number 3-0000-12435 from Infect-ERA and the Chief Scientist’s Bureau in the Israeli Ministry of Health.

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Copyright information

© Springer Science+Business Media, LLC 2018

Authors and Affiliations

  1. 1.The Infectious Diseases Research LaboratorySheba Medical CenterTel-Hashomer, Ramat GanIsrael
  2. 2.Department of Clinical Microbiology and Immunology, Sackler Faculty of MedicineTel Aviv UniversityTel AvivIsrael

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