Development of a dual promoter-mediated gene silencing system in Flammulina velutipes
Development of a dual promoter-mediated gene silencing system in Flammulina velutipes
复制标题
DOI：
10.1016/j.myc.2017.01.003
复制
发表时间：
2017-05
期刊：
影响因子：
1.4
通讯作者：
Liang Shi;Tian-Jun Zhang;Chao Xu;Ang Ren;Ailiang Jiang;Han-shou Yu;Mingwen Zhao
中科院分区：
文献类型：
--
作者：
Liang Shi;Tian-Jun Zhang;Chao Xu;Ang Ren;Ailiang Jiang;Han-shou Yu;Mingwen Zhao
本研究旨在建立一个基于双启动子构建的RNAi系统，以干扰金针菇中目的基因的表达。本研究将克隆的内源漆酶基因作为沉默报告基因导入F。通过电穿孔介导的转化将其转化到金针菇中。我们的数据明确表明，双启动子沉默系统显着降低lac 6的表达和活性。此外，lac 6 mRNA水平和酶活性的降低是相关的（至少降低50%）。本研究中开发的分子工具应有助于这一重要物种基因的功能表征。© 2017日本真菌学会。由爱思唯尔BV出版版权所有。不包括法国）唱歌是世界各地栽培的著名食用菌。它是世界上最活跃的六种蘑菇之一;每年生产超过30万吨这种蘑菇（Kim等人，2014）。近年来，许多研究报道了F. Velutipes，如其抗氧化和免疫调节活性（Zhang et al. 2013; Wu et al. 2014; Ma et al. 2015）。近年来，对F.一些研究包括基因组序列分析（Kim et al. 2014; Park et al. 2014; Kurata et al. 2016）和基因功能分析（Nakai et al. 2000; Kuo et al. 2004; Yamada et al. 2005，2008）。本文主要研究了黄曲霉的生长发育和代谢规律。尽管已经进行了一些基因转移程序，但velutipes仍处于早期阶段。
This study aimed to establish an RNAi system based on a dual promoter construct to interfere with the expression of target genes in Flammulina velutipes. In this study, the endogenous laccase gene, which was cloned as a silencing reporter, was introduced into F. velutipes through electroporation-mediated transformation. Our data unequivocally indicate that the dual promoter silencing system significantly reduced the expression and activity of lac6. Additionally, reductions in lac6 mRNA levels and enzyme activity were correlated (minimum of a 50% reduction). The molecular tools developed in this study should facilitate the functional characterization of genes in this important species.© 2017 The Mycological Society of Japan. Published by Elsevier BV All rights reserved.Flammulina velutipes (Curt. ex Fr.) Sing. is a well-known edible mushroom cultivated all over the world. It is one of the six most actively cultivated mushrooms in the world; over 300,000 tons of this mushroom are produced per year (Kim et al. 2014). For the past several years, many studies have described the important biological activity of F. velutipes, such as its antioxidant and immunomodulatory activities (Zhang et al. 2013; Wu et al. 2014; Ma et al. 2015). The number of studies investigating the molecular genetics of F. velutipes has increased, with some studies including genome sequence analysis (Kim et al. 2014; Park et al. 2014; Kurata et al. 2016) and gene function analysis (Nakai et al. 2000; Kuo et al. 2004; Yamada et al. 2005, 2008). The research on the development and metabolism of F. velutipes is still at an early stage, although some gene transfer procedures have been