Journal: Genes

Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape

doi: 10.3390/genes15111478

Figure Lengend Snippet: Gene structure and amino acid sequence alignment analysis of VvCYP76F14 proteins. ( A ) Gene structure of VvCYP76F14 s derived from three mutant lines. ( B ) Acid sequence alignment of VvCYP76F14 proteins.

Article Snippet: Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 .

Techniques: Sequencing, Derivative Assay, Mutagenesis

Journal: Genes

Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape

doi: 10.3390/genes15111478

Figure Lengend Snippet: Subcellular localization of VvCYP76F14. The CDS of VvCYP76F14 derived from ‘Yantai 2-2-08’ and ‘Yantai 2-2-08’ and further cloned into the pBWA(V)HS-ccdb-GLosgfp vector. The GV3101 strain harboring the pBWA(V)HS-CYP76F14-Glosgfp or the empty vector was infiltrated into Arabidopsis mesophyll protoplasts. The GFP fluorescence and chloroplast autofluorescence were observed using the excitation/emission wavelengths 470/510 nm and 620/660 nm, respectively. Scale bar = 10 μm.

Article Snippet: Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 .

Techniques: Derivative Assay, Clone Assay, Plasmid Preparation, Fluorescence

Journal: Genes

Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape

doi: 10.3390/genes15111478

Figure Lengend Snippet: In vitro activity analysis of VvCYP76F14 in Escherichia coli . The in vitro enzymatic activity of VvCYP76F14 was assayed by measuring the amount of remnant substrate. Data are shown as means ± SE ( n = 3). Letters indicate significant differences among five VvCYP76F14s at a significance level of p ≤ 0.05, as determined using ANOVA followed by Fisher’s LSD test.

Article Snippet: Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 .

Techniques: In Vitro, Activity Assay

Journal: Genes

Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape

doi: 10.3390/genes15111478

Figure Lengend Snippet: Transient expression of VvCYP76F14 in berries of D299T mutant lines. The content levels of linalool, ( E )-8-hydroxylinalool, ( E )-8-oxolinalool, and ( E )-8-carboxylinalool in berries of ‘Yantai 2-2-08’ ( A ), ‘Yantai 2-2-19’ ( B ), and ‘Yantai 2-3-37’ ( C ) were assayed by UPLC-MS. Data are shown as means ± SE ( n = 3). Letters indicate significant differences among five VvCYP76F14s at a significance level of p ≤ 0.05, as determined using ANOVA followed by Fisher’s LSD test.

Article Snippet: Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 .

Techniques: Expressing, Mutagenesis