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plant expression vector pmdc32 hpb  (Addgene inc)


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    Addgene inc plant expression vector pmdc32 hpb
    Plant Expression Vector Pmdc32 Hpb, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/pmdc32+hpb/pMDC32-HPB+(Plasmid+%2332078)/bio_rxiv__2025__05__06__652549-219-13-18
    Average 93 stars, based on 13 article reviews
    plant expression vector pmdc32 hpb - by Bioz Stars, 2026-09
    93/100 stars

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    other:

    Article Title: Differences in PpAAT1 Activity in High- and Low-Aroma Peach Varieties Affect γ -Decalactone Production
    Article Snippet: For the MS/MS detection, all precursors were fragmented using 20 to 40 eV, and the scan time was 0.2 s. The MS data were analyzed using the Progenesis QI program (version 2.2; Waters).

    Expressing:

    Article Title: Differences in PpAAT1 Activity in High- and Low-Aroma Peach Varieties Affect γ -Decalactone Production
    Article Snippet: A constant flow rate of 0.4 mL min −1 was used with the following gradient elution conditions: initially 100% A, held for 0 to 2 min; then 2 to 9 min 0% B to 100% B; and then 100% B, held for 9 to 11 min. A high-resolution tandem mass spectrometer (Xevo G2-XS QTOF; Waters) was used to detect the metabolites eluted from the column. .. Transient Expression in Nicotiana benthamiana The PpAAT1 cDNA clone was transferred into the pMDC32-HPB (Addgene no. 32078) plasmid vector using the Gateway LR Clonase II Enzyme mix (Thermo Fisher Scientific) and transformed into Agrobacterium tumefaciens LBA4404 Electro-Cells (Takara) according to the manufacturer’s instructions. .. For the MS/MS detection, all precursors were fragmented using 20 to 40 eV, and the scan time was 0.2 s. The MS data were analyzed using the Progenesis QI program (version 2.2; Waters).

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Article Title: Potential role of fatty acid desaturase 2 in regulating peach aroma formation
    Article Snippet: Fruit aroma is an important evaluation index for fruit quality.. Linoleic acid is an essential precursor pool for the aroma compounds in peach (Prunus persica L.) and is catalyzed by fatty acid desaturase 2 (FAD2).. In this study, PpFAD2–1 and PpFAD2–2 were isolated from ‘Jinxiuhuangtao’ peaches.

    Plasmid Preparation:

    Article Title: Differences in PpAAT1 Activity in High- and Low-Aroma Peach Varieties Affect γ -Decalactone Production
    Article Snippet: A constant flow rate of 0.4 mL min −1 was used with the following gradient elution conditions: initially 100% A, held for 0 to 2 min; then 2 to 9 min 0% B to 100% B; and then 100% B, held for 9 to 11 min. A high-resolution tandem mass spectrometer (Xevo G2-XS QTOF; Waters) was used to detect the metabolites eluted from the column. .. Transient Expression in Nicotiana benthamiana The PpAAT1 cDNA clone was transferred into the pMDC32-HPB (Addgene no. 32078) plasmid vector using the Gateway LR Clonase II Enzyme mix (Thermo Fisher Scientific) and transformed into Agrobacterium tumefaciens LBA4404 Electro-Cells (Takara) according to the manufacturer’s instructions. .. For the MS/MS detection, all precursors were fragmented using 20 to 40 eV, and the scan time was 0.2 s. The MS data were analyzed using the Progenesis QI program (version 2.2; Waters).

    Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape
    Article Snippet: To evaluate the enzymatic activity of VvCYP76F14 in berries, the CDS of VvCYP76F14 was isolated from ‘L35’, ‘Merlot’, or D299T mutant lines, respectively, and synthesized by GenScript Co., Ltd. (Nanjing, China). .. Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 . ..

    Transformation Assay:

    Article Title: Differences in PpAAT1 Activity in High- and Low-Aroma Peach Varieties Affect γ -Decalactone Production
    Article Snippet: A constant flow rate of 0.4 mL min −1 was used with the following gradient elution conditions: initially 100% A, held for 0 to 2 min; then 2 to 9 min 0% B to 100% B; and then 100% B, held for 9 to 11 min. A high-resolution tandem mass spectrometer (Xevo G2-XS QTOF; Waters) was used to detect the metabolites eluted from the column. .. Transient Expression in Nicotiana benthamiana The PpAAT1 cDNA clone was transferred into the pMDC32-HPB (Addgene no. 32078) plasmid vector using the Gateway LR Clonase II Enzyme mix (Thermo Fisher Scientific) and transformed into Agrobacterium tumefaciens LBA4404 Electro-Cells (Takara) according to the manufacturer’s instructions. .. For the MS/MS detection, all precursors were fragmented using 20 to 40 eV, and the scan time was 0.2 s. The MS data were analyzed using the Progenesis QI program (version 2.2; Waters).

    Activity Assay:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Isolation:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Mutagenesis:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Synthesized:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Clone Assay:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape
    Article Snippet: To evaluate the enzymatic activity of VvCYP76F14 in berries, the CDS of VvCYP76F14 was isolated from ‘L35’, ‘Merlot’, or D299T mutant lines, respectively, and synthesized by GenScript Co., Ltd. (Nanjing, China). .. Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 . ..

    Virus:

    Article Title: Site-directed mutagenesis identified the key active site residues of alcohol acyltransferase PpAAT1 responsible for aroma biosynthesis in peach fruits
    Article Snippet: The turnover number ( k cat ) and affinity ( K m ) were calculated using Origin 2018 ( www.originlab.com ). .. To assess the enzymatic activity of PpAAT1 and PpAAT1-SMs in plants, PpAAT1 isolated from ‘Fenghuayulu’ and its mutant forms ( PpAAT1 - SMs ) synthesized at GenScript Co. Ltd. (Nanjing, China) were cloned into pMDC32-HPB (Addgene: 32078), driven by the cauliflower mosaic virus 35S promoter, to generate the plant expression vectors pMDC32-HPB- PpAAT1 and pMDC32-HPB- PpAAT1 - SMs . .. To assess the internal esterification activity of PpAAT1 and PpAAT1-SMs in tobacco, the GV3101 (pSoup-P19) (Shanghai Weidi Biotechnology Co., Ltd) strain containing pMDC32-HPB- PpAAT1 or each of the 14 pMDC32-HPB- PpAAT1 - SMs was infiltrated into tobacco leaves, together with the substrate of 4-hydroxy-decanoyl–CoA (10 mM), as described previously .

    Amplification:

    Article Title: Potential role of fatty acid desaturase 2 in regulating peach aroma formation
    Article Snippet: Fruit aroma is an important evaluation index for fruit quality.. Linoleic acid is an essential precursor pool for the aroma compounds in peach (Prunus persica L.) and is catalyzed by fatty acid desaturase 2 (FAD2).. In this study, PpFAD2–1 and PpFAD2–2 were isolated from ‘Jinxiuhuangtao’ peaches.

    Sequencing:

    Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape
    Article Snippet: To evaluate the enzymatic activity of VvCYP76F14 in berries, the CDS of VvCYP76F14 was isolated from ‘L35’, ‘Merlot’, or D299T mutant lines, respectively, and synthesized by GenScript Co., Ltd. (Nanjing, China). .. Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 . ..

    Recombinant:

    Article Title: D299T Mutation in CYP76F14 Led to a Decrease in Wine Bouquet Precursor Production in Wine Grape
    Article Snippet: To evaluate the enzymatic activity of VvCYP76F14 in berries, the CDS of VvCYP76F14 was isolated from ‘L35’, ‘Merlot’, or D299T mutant lines, respectively, and synthesized by GenScript Co., Ltd. (Nanjing, China). .. Sequencing-verified VvCYP76F14 was cloned into the pMDC32-HPB (Addgene: 32078) to generate the recombinant plasmid pMDC32-HPB- CYP76F14 . ..



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    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.

    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

    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.

    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

    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.

    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

    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.

    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