【摘要】：目的篩選效率最高的酵母前體飼喂方式,為在酵母中進(jìn)行基因功能驗(yàn)證提供方法學(xué)指導(dǎo)。方法首先構(gòu)建含PsP6H基因和PsCPR基因重組酵母工程菌,然后用4種不同的方法進(jìn)行前體飼喂,最后質(zhì)譜檢測下游產(chǎn)物二氫血根堿來確定最優(yōu)方案。結(jié)果最優(yōu)的前體飼喂方法是將酵母工程菌經(jīng)過半乳糖誘導(dǎo)31 h后在pH 8.0的Tris EDTA溶液中孵育24 h。結(jié)論不同的前體飼喂方式對酵母工程菌下游產(chǎn)物產(chǎn)生不同影響,經(jīng)優(yōu)化后的酵母前體飼喂方法較穩(wěn)定,重復(fù)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.
【作者單位】： 湖南農(nóng)業(yè)大學(xué)園藝園林學(xué)院;湖南農(nóng)業(yè)大學(xué)獸用中藥資源與中獸藥創(chuàng)制國家地方聯(lián)合工程研究中心;湖南中醫(yī)藥大學(xué)藥學(xué)院;湖南農(nóng)業(yè)大學(xué)動物醫(yī)學(xué)院;
【基金】：國家重點(diǎn)實(shí)驗(yàn)室培育基地開放項(xiàng)目(15KFXM15) 園藝學(xué)優(yōu)勢特色重點(diǎn)學(xué)科開放基金項(xiàng)目(2015YYX001) 湖南農(nóng)業(yè)大學(xué)人才引進(jìn)基金(13YJ09) 湖南省研究生科研創(chuàng)新項(xiàng)目(CX2014B302)
【分類號】：Q78
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本文編號：2307727