alexa Abstract | 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
Open Access

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Research Article Open Access


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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Author(s): Haizhao Xue, Yibo Hu, Xiaoran Fang,Guangshan Yao, Xin Song

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