酵母破壁酶和蜗牛酶辅助TRIzol试剂法提取酿酒酵母总RNA  

Total RNA extraction from Saccharomyces cerevisiae by yeast lyticase and snailase assisted TRIzol reagent method

作　　者：安佳星 伍时华[1,2,3] 黄嘉齐 曾令杰 黄锦翔 丰丕雪 易弋[1,2,3] AN Jiaxing;WU Shihua;HUANG Jiaqi;ZENG Lingjie;HUANG Jinxiang;FENG Pixue;YI Yi(College of Biological and Chemical Engineering,Guangxi University of Science and Technology,Liuzhou 545006,China;Guangxi Key Laboratory of Green Processing of Sugar Resources,Guangxi University of Science and Technology,Liuzhou 545006,China;Key Laboratory for Processing of Sugar Resources of Guangxi Higher Education Institutes,Guangxi University of Science and Technology,Liuzhou 545006,China)

机构地区：[1]广西科技大学生物与化学工程学院,广西柳州545006 [2]广西科技大学广西糖资源绿色加工重点实验室,广西柳州545006 [3]广西科技大学广西高校糖资源加工重点实验室,广西柳州545006

出　　处：《中国酿造》2020年第12期46-50,共5页China Brewing

基　　金：国家自然科学基金(No.31560728);广西自然科学基金(No.2018GXNSFAA050116)。

摘　　要：该研究以酵母破壁酶、蜗牛酶破碎酿酒酵母(Saccharomyces cerevisiae)细胞壁,再使用总核糖核酸(RNA)提取试剂(TRIzol)法提取酵母总RNA,通过对总RNA进行浓度与纯度的分析,比较酶处理对RNA提取的影响。结果表明,酵母破壁酶的最佳提取条件为加酶量37.5μL,提取温度27℃,提取时间15 min。在此最佳提取条件下,提取RNA质量浓度为1079.05 ng/μL,A260 nm/A280 nm为2.10,A260 nm/A230 nm为2.15;蜗牛酶的最佳提取条件为加酶量30 mg/mL,提取温度37℃,提取时间30 min。在此最佳提取条件下,提取RNA质量浓度为1673.39 ng/μL,A260 nm/A280 nm为1.99,A260 nm/A230 nm为1.81。结果显示,酵母破壁酶的提取效果优于蜗牛酶。The cell wall of Saccharomyces cerevisiae was broken using yeast lyticase and snailase,and then the total ribonucleic acid(RNA)of yeast was extracted by total RNA extraction reagent(TRIzol)method.Through the analysis of the concentration and purity of the total RNA,the effect of enzyme treatment on RNA extraction was compared.The results showed that the optimal extraction conditions of yeast lyticase were enzyme addition 37.5μl,extraction temperature 27℃,and extraction time 15 min.Under the optimal extraction conditions,the mass concentration of RNA was 1079.05 ng/μl,A260 nm/A280 nm was 2.10,and A260 nm/A230 nm was 2.15.The optimal extraction conditions of snailase were enzyme addition 30 mg/ml,extraction temperature 37℃,and extraction time 30 min.Under the optimal extraction conditions,the mass concentration of RNA was 1673.39 ng/μl,A260 nm/A280 nm was 1.99,and A260 nm/A230 nm was 1.81.At the same time,it was found that the extraction effect of yeast lyticase was better than that of snailase.

关 键 词：TRIzol试剂法 蜗牛酶 酵母破壁酶 酿酒酵母 RNA提取 

分 类 号：Q939.5[生物学—微生物学]