期刊
MOLECULAR BIOTECHNOLOGY
卷 35, 期 3, 页码 283-294出版社
HUMANA PRESS INC
DOI: 10.1007/BF02686014
关键词
RNAi hairpin; post-transcriptional gene-silencing (PTGS); Coprinus cinereus; green fluorescent protein (GFP); silencing cassettes; antisense; Basidiomycetes; Agaricus bisporus
Post-transcriptional gene-silencing methods (PTGS), including RNAi, are becoming pervasive in functional genomics. To advance analysis of the recently sequenced Coprinus cinereus genome, a high throughput gene silencing method is essential. We have exploited the GFP reporter gene to evaluate and quantify efficacy of three different silencing strategies. Modular constructs that encompassed antisense, untranslatable sense, and RNAi-mediating hairpin sequences, were transformed into a GFP-expressing host strain. Transformants exhibiting strong downregulation and partial suppression of GFP were recovered with all three constructs. Analyses of protein and transcriptional nucleic acids revealed that the antosense and hairpin sequences yielded similar levels of GFP suppression, and were both more efficient than untranslatable sense sequence. Our antisense vectors will expedite functional characterisation of C. cinereus and the modular nature of the constructs should permit exploitation of directional cDNA libraries for high throughput screening.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据