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Development of orphan G-protein coupled receptor peptidic ligand identification system

 

 

Project Title: „Development of orphan G-protein coupled receptor peptidic ligand identification system” 

Funding: European Regional Development Fund (ERDF), Measure 1.1.1.1 “Industry-Driven Research”

Project Nr.: 1.1.1.1/16/A/055

Period: 1st January 2017 – 31st December 2019

Project costs: 285 245,84 EUR

Principle Investigator: Dr. biol. D. Fridmanis

 

The aim of this Project is to develop humanized yeast S.cerevisiae based GPCR peptide ligand identification system and to evaluate its effectiveness by de-orphanization of C.intestinalis orexin-like receptor and determination of novel ligand’s pharmacological properties. The results of this Project shall provide instrument for the development of novel pharmaceutical compounds and promote development of biopharmaceutical industry in Latvia.

Information published: 02.01.2017.

 

Progress of the project

1 January 2017 - 31 March 2017

Experimental development

According to plan the first three months of the project were devoted to creation of double expression plasmid. Since p426GPD and p426TEF expression plasmids that have been successfully employed by our group are nearly identical, then within the scope of these activities we altered the multi-clonal sites of the expression loci in a way that each would contain only two unique cloning/restriction sites, thus becoming mutually compatible. Following the modification of both plasmids we performed the sequencing analysis of both expression loci to ascertain the conformity of acquired sequences to planned ones and carried out the transfer of TEF containing locus from p426TEF to p426GPD plasmid. At the moment we are performing the sequencing analysis of thus acquired construct.

Information published: 31.03.2017.


Progress of the project

1 April 2017 - 30 June 2017

Experimental development

During this project phase the functional regions of TEF promoter containing plasmid that were relevant for gene expression were transferred to GDP promoter containing plasmid. Further within thus acquired double expression plasmid MC4R and a-MSH coding genes were inserted and following their insertion conformity of acquired results with the plans was established by sequencing analysis. After acquisition of expression constructs we commenced with evaluation of its functionality.

Information published: 30.06.2017.

 

Progress of the project

1 July 2017 - 30 September 2017

Experimental development

During this project phase we initiated the functional testing of previously created double expression plasmid. Within the scope of these activities the newly created plasmid was inserted into the cells of humanized yeast S.cerevisiae strain MMY28, which were subsequently seeded within the selective media containing petri dish. The media within selected dishes was supplemented with MC4R ligand - a-MSH as the means of positive control. Acquired results were ambiguous therefore experiments shall be repeated during further project stages.

Information published: 29.09.2017.



Mājas lapas izstrādi finansēja ERAF 2.1.1.2. aktivitātes projekts Nr. 2010/0196/2DP/2.1.1.2.0/10/APIA/VIAA/004 "Latvijas biomedicīnas pētījumu integrācija Eiropas zinātnes telpā".