High Efficiency and Stable RNA Interference Vector Construction for Penicillium sp.
- *Corresponding Author:
- Xin Song
State Key Laboratory of Microbial Technology
School of Life Science, Shandong University
27 Shandanan Road, Jinan,
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
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.