alexa High Efficiency and Stable RNA Interference Vector Construction for Penicillium sp.
E- ISSN: 2320 - 3528
P- ISSN: 2347 - 2286

Research & Reviews: Journal of Microbiology and Biotechnology
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Research Article

High Efficiency and Stable RNA Interference Vector Construction for Penicillium sp.

Haizhao Xue1, Yibo Hu1, Xiaoran Fang1, Guangshan Yao1 Xin Song1,2*

1State Key Laboratory of Microbial Technology, School of Life Science, Shandong University, Shan Da Nan Road 27, Jinan 250100, China

2National Glycoengineering Research Center, Shandong University, Shan Da Nan Road 27, Jinan 250100, Shandong, People’s Republic of China

*Corresponding Author:
Xin Song
State Key Laboratory of Microbial Technology
School of Life Science, Shandong University
27 Shandanan Road, Jinan,
Shandong 250100
P. R. China
Tel: +86 53188565610
E-mail: [email protected]

Received date: 30/11/2016; Accepted date: 28/06/2016; Published date: 06/07/2016

 

Abstract

RNA interference (RNAi), which can turn down gene expression effectively and specifically, has showed great potential in gene function research and gene modification. In this study, a RNA interference vector pHX-RNAi, which carries a hygromycin resistance cassette and a transcriptional unit for hairpin RNA expression was constructed. A gene amy15A which encodes the major amylase in a high cellulase producer strain Penicillium oxalicum A11 was targeted knocking-down. The results of SDS-PAGE and RT-qPCR showed that both the transcription and secretion levels of Amy15A were dramatically down-regulated, which implying that RNAi play a significant role in regulation of gene expression post-transcriptionally. All of the hygromycin-resistant transformants with the silencing constructs showed high degree of silencing in expression of amy15A. Considering the conservatism of RNAi machinery in fungi, the RNAi vector established in this study- pHX-RNAi could be applied to other filamentous fungi.

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