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. 2016 Jun 27;6:28541. doi: 10.1038/srep28541

Figure 3. Functional expression of GmDGATs in yeast cells.

Figure 3

The quadruple mutant Saccharomyces cerevisiae strain H1246 and the wild-type strain YPH499a were transformed with GmDGAT genes and vector control. (a) Nile red staining of oil drops in GmDGAT1A (i, vi)- and 2D (iii, viii)- or pYESDEST52 vector (ii, vii)-expressing yeast mutant strain H1246, and vector (iv, ix) and GmDGAT2D (v, x expressed in wild-type strain YPH499a cells. TAG was extracted and separated by thin-layer chromatography and TAG spots were scraped for analyzing compositions of fatty acids by GC. (b) Nile Red Assay on oil drops and fluorescence-based TAG content. (c) Thin-layer chromatography of lipids extracted from yeast transformants. DAG, fatty acids (FAs), and TAG spots were labeled out. TAG spots were scraped off from the thin-layer chromatography (TLC) plates and were extracted for analysis of TAG contents and compositions with GC. (d) Total TAG production in yeast mutant strain H1246 and wild-type strain YPH499a, Total TAG was measured by GC. (e) Fatty acid compositions of TAG produced in GmDGAT2D- or pYESDEST52 empty vector-expressing yeast mutant strain H1246 and wild-type strain YPH499a. All data are presented by mean of at least three biological replicates ± SD. *P < 0.05 and **P < 0.01 by Student’s t test for significant difference.