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

Haizhao Xue, Yibo Hu, Xiaoran Fang,Guangshan Yao, Xin Song

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.