Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Oct 21;12:17658. doi: 10.1038/s41598-022-21678-3

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

PMC Copyright notice

Construction of multigene expression vectors and biochemical analysis. (a) Assembly of individual expression vectors encoding TAS2R, mt-clytin II, and Gα16-gust44 by in vitro Cre-loxP recombination to generate type 1 and type 2 multigene vectors. With TAS2R as the gene of reference, the expression vectors are arranged in the following sequences in clockwise fashion: TAS2R–mt-clytin II–Gα16-gust44 (type 1) and TAS2R–Gα16-gust44–mt-clytin II (type 2). In both type 1 and type 2 plasmids, TAS2R is divergently orientated from mt-clytin II and Gα16-gust44 chimera. Black arrows indicate the direction of expression from the CMV or CAG promoter. Grey circles represent loxP sites. Only relevant elements of the expression constructs are shown for figure clarity. (b) Concentration–response curves of TAS2Rs upon stimulation with their cognate agonists in the bioluminescence-based intracellular calcium release assay. Type 2 vectors consistently produced larger assay spans than that of type 1 vectors. Data points are shown as mean ± s.e.m. from a representative experiment out of three independent biological replicates performed in technical quadruplicates. EC₅₀, half-maximal effective concentration. Figure 1a was generated using Adobe Illustrator (Version 25.4.1).