NDLI: Temporal Quantification of MAPK Induced Expression in Single Yeast Cells

Content Provider	PubMed Central
Author	Pelet, Serge Aymoz, Delphine Eric, Durandau
Copyright Year	2013
Abstract	The quantification of gene expression at the single cell level uncovers novel regulatory mechanisms obscured in measurements performed at the population level. Two methods based on microscopy and flow cytometry are presented to demonstrate how such data can be acquired. The expression of a fluorescent reporter induced upon activation of the high osmolarity glycerol MAPK pathway in yeast is used as an example. The specific advantages of each method are highlighted. Flow cytometry measures a large number of cells (10,000) and provides a direct measure of the dynamics of protein expression independent of the slow maturation kinetics of the fluorescent protein. Imaging of living cells by microscopy is by contrast limited to the measurement of the matured form of the reporter in fewer cells. However, the data sets generated by this technique can be extremely rich thanks to the combinations of multiple reporters and to the spatial and temporal information obtained from individual cells. The combination of these two measurement methods can deliver new insights on the regulation of protein expression by signaling pathways.
Related Links	http://dx.doi.org/10.3791/50637
Starting Page	50637
File Format	PDF
ISSN	1940087X
e-ISSN	1940087X
Journal	Journal of Visualized Experiments : JoVE
Issue Number	80
Language	English
Publisher	MyJove Corporation
Publisher Date	2013-10-01
Access Restriction	Open
Rights Holder	MyJove Corporation
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Immunology and Microbiology(all) Chemical Engineering(all) Neuroscience(all) Medicine(all) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Neuroscience Immunology and Microbiology Biochemistry, Genetics and Molecular Biology Chemical Engineering

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Identification of Growth Inhibition Phenotypes Induced by Expression of Bacterial Type III Effectors in Yeast

Purification of Mitochondria from Yeast Cells

Transcriptome Analysis of Single Cells

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

A Rapid Technique for the Visualization of Live Immobilized Yeast Cells

Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 1

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 2

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 3

Temporal Quantification of MAPK Induced Expression in Single Yeast Cells

Similar Documents

Identification of Growth Inhibition Phenotypes Induced by Expression of Bacterial Type III Effectors in Yeast

Purification of Mitochondria from Yeast Cells

Transcriptome Analysis of Single Cells

Expression of Recombinant Proteins in the Methylotrophic Yeast Pichia pastoris

A Rapid Technique for the Visualization of Live Immobilized Yeast Cells

Quantifying Yeast Chronological Life Span by Outgrowth of Aged Cells

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 1

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 2

Vaccinia Virus Infection & Temporal Analysis of Virus Gene Expression: Part 3

Temporal Quantification of MAPK Induced Expression in Single Yeast Cells