贝莱斯芽孢杆菌总RNA提取的Trizol改良法  

A Modified Trizol Method for Total RNA Isolation from Bacillusvelezensis

作　　者：王松 龚禹瑞 袁亚男 尚静静 张树竹 陈本佳 秦世雯 WANG Song;GONG Yurui;YUAN Ya'nan;SHANG Jingjing;ZHANG Shuzhu;CHEN Benjia;QIN Shiwen(School of Agriculture,Yunnan University/Yunnan Engineering and Technology Research Center of Perennial Rice,Kunming,Yunnan 650500,China)

机构地区：[1]云南大学农学院/云南省多年生稻工程技术研究中心,云南昆明650500

出　　处：《热带农业科学》2020年第7期52-58,共7页Chinese Journal of Tropical Agriculture

基　　金：云南省科技厅科技计划项目(No.2019FD128);云南省教育厅科学研究基金(No.2018JS007)。

摘　　要：贝莱斯芽孢杆菌(Bacillus velezensis)是在农业、环境和发酵工业等领域具有重要应用潜力的细菌。但芽孢杆菌属细菌具有较厚的细胞壁且分泌蛋白多,常规方法难以抽提到高质量的总RNA。因此,建立和优化简便有效的贝莱斯芽孢杆菌总RNA提取方法,将为B.velezensis基因表达和功能研究提供重要的技术支撑。通过优化传统Trizol法,对菌体培养条件、菌体洗涤次数、液氮冷冻处理和溶菌酶裂解菌体方法进行改良,建立了一种简便有效的B.velezensis总RNA提取方法。Bacillus velezensis is a Gram-positive bacteria,and it has prospects of application in agriculture,environment and fermentation industry.However,it is difficult to extract RNA with high quality and quantity from Bacillus spp.,because they have thick cell wall and secrete plenty of proteins that are easy to result in contamination.Therefore,a simple and effective method for total RNA isolation could provide technical support for the gene expression and gene function analysis of B.velezensis.A Trizol-based RNA extraction method for B.velezensis was optimized by modifying the bacterial cell growth conditions,washing times,liquid nitrogen freezing treatment,and lysozymatic break down of the bacterial cell wall,and a simple and effective method for total RNA isolation of B.velezensis was hence established.

关 键 词：贝莱斯芽孢杆菌 总RNA提取 Trizol改良法 

分 类 号：S432.1[农业科学—植物病理学]