酵母前体饲喂验证植物次生代谢产物功能基因方法学初探
发布时间：2018-11-03 12:02
【摘要】：目的筛选效率最高的酵母前体饲喂方式,为在酵母中进行基因功能验证提供方法学指导。方法首先构建含PsP6H基因和PsCPR基因重组酵母工程菌,然后用4种不同的方法进行前体饲喂,最后质谱检测下游产物二氢血根碱来确定最优方案。结果最优的前体饲喂方法是将酵母工程菌经过半乳糖诱导31 h后在pH 8.0的Tris EDTA溶液中孵育24 h。结论不同的前体饲喂方式对酵母工程菌下游产物产生不同影响,经优化后的酵母前体饲喂方法较稳定,重复性好。
[Abstract]:Objective to screen the most efficient yeast precursor feeding method and to provide methodological guidance for gene function verification in yeast. Methods Recombinant yeast engineering strains containing PsP6H gene and PsCPR gene were first constructed, and then were fed with four different methods. Finally, the downstream product dihydrohaematine was detected by mass spectrometry to determine the optimal scheme. Results the optimal precursor feeding method was to incubate the engineered yeast strain in Tris EDTA solution of pH 8.0 for 24 h after induction with galactose for 31 h. Conclusion different methods of precursor feeding have different effects on the downstream products of yeast engineering bacteria. The optimized method of yeast precursor feeding is stable and reproducible.
【作者单位】： 湖南农业大学园艺园林学院;湖南农业大学兽用中药资源与中兽药创制国家地方联合工程研究中心;湖南中医药大学药学院;湖南农业大学动物医学院;
【基金】：国家重点实验室培育基地开放项目(15KFXM15) 园艺学优势特色重点学科开放基金项目(2015YYX001) 湖南农业大学人才引进基金(13YJ09) 湖南省研究生科研创新项目(CX2014B302)
【分类号】：Q78
，
本文编号：2307727
[Abstract]:Objective to screen the most efficient yeast precursor feeding method and to provide methodological guidance for gene function verification in yeast. Methods Recombinant yeast engineering strains containing PsP6H gene and PsCPR gene were first constructed, and then were fed with four different methods. Finally, the downstream product dihydrohaematine was detected by mass spectrometry to determine the optimal scheme. Results the optimal precursor feeding method was to incubate the engineered yeast strain in Tris EDTA solution of pH 8.0 for 24 h after induction with galactose for 31 h. Conclusion different methods of precursor feeding have different effects on the downstream products of yeast engineering bacteria. The optimized method of yeast precursor feeding is stable and reproducible.
【作者单位】： 湖南农业大学园艺园林学院;湖南农业大学兽用中药资源与中兽药创制国家地方联合工程研究中心;湖南中医药大学药学院;湖南农业大学动物医学院;
【基金】：国家重点实验室培育基地开放项目(15KFXM15) 园艺学优势特色重点学科开放基金项目(2015YYX001) 湖南农业大学人才引进基金(13YJ09) 湖南省研究生科研创新项目(CX2014B302)
【分类号】：Q78
，
本文编号：2307727
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