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. 2015 Aug 11;12(8):824–837. doi: 10.1080/15476286.2015.1058476

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

© Georis Isabelle, Jennifer J Tate, Fabienne Vierendeels, Terrance G Cooper, and Evelyne Dubois

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 4. — Two mRNAs are produced from the GAT1 locus. Total RNA was isolated from wild type (TB50), gat1Δ (FV006) and GAT1-MYC¹³ cells of the TB (FV063), FY (FV034) and Sigma (FV291) genetic backgrounds that were grown in YNB medium with glutamine (Gln), proline (Pro) or ammonium (Am.) as the nitrogen source and treated with methionine sulfoximine (Am. + Msx). 30 μg of total RNA from each sample were subjected to Northern blot analysis. HHT1 was used as the loading and transfer efficiency control. (A). GAT1 mRNA analysis in wild type GAT1-MYC¹³ cells using a double stranded probe covering the 5′ region of the gene (GAT1_O7-GAT1_O10). (B). GAT1 mRNA analysis in wild type GAT1-MYC¹³ cells using a double stranded probe covering the 3′ region of the gene (GAT1_O15-GAT1_O4). (C). GAT1 mRNA analysis in wild type untagged and GAT1-MYC¹³ cells from 3 different genetic backgrounds using a double stranded probe covering the 5′ region of the gene (GAT1_O7-GAT1_O10). (D). Wild type cells of Saccharomyces paradoxus, bayanus and mikatae were grown in YNB medium with glutamine (Gln) or proline (Pro) as the nitrogen source. Total RNA was isolated and GAT1 mRNA levels were assessed using qRTPCR as described in Figure 3 using GAT1_O1-O2 and GAT1_O3-O4 primer pairs specific for each species. (E). GAT1 mRNA analysis in wild type untagged (TB50), gat1Δ (FV006) and GAT1-MYC¹³ (FV063) cells using strand-specific probes covering the 5′ region of the gene (GAT1_O7-GAT1_O10).