NDLI: The first reporter gene assay on living cells : Green fluorescent protein as reporter gene for the investigation of Gi-protein coupled receptors

Content Provider	Springer Nature Link
Author	Dinger, Michaela C. Beck Sickinger, Annette G.
Copyright Year	2002
Abstract	Reporter gene assay systems are important tools for the investigation of G-protein coupled receptors and their interaction with ligands. Here, we describe a novel reporter gene assay system for the investigation of G$_{i}$-protein coupled receptors in living cells. For the first time green fluorescent protein (GFP) was used as reporter gene under the transcriptional control of cAMP-response elements (CREs). Three different reporter gene vectors with increasing numbers of CREs were cloned and GFP expression was investigated after forskolin stimulation. Furthermore, the novel reporter system was successfully applied to the neuropeptide Y (NPY) rY$_{5}$ receptor subtype, a G$_{i}$-protein coupled receptor. Our data clearly demonstrate dose-dependent GFP expression for NPY. Furthermore, receptor subtype selectivity of well characterized NPY analog could be proven for the NPY rY$_{5}$ receptor. The great advantage of the method is that no cell lysis is required and assays can be performed on living cells. Accordingly, in vitro testing of agonist gets faster and significantly more convenient.
Starting Page	9
Ending Page	18
Page Count	10
File Format	PDF
ISSN	10736085
Journal	Molecular Biotechnology
Volume Number	21
Issue Number	1
e-ISSN	15590305
Language	English
Publisher	Humana Press
Publisher Date	2002-01-01
Publisher Place	Totowa
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Neurosciences Neurology
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biochemistry Bioengineering Applied Microbiology and Biotechnology Biotechnology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
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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
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