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Quantifiable downregulation of endogenous genes in Agaricus bisporus mediated by expression of RNA hairpins

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Costa, Ana M. S. B., Thomas, D. John I., Eastwood, Daniel C., Cutler, Simon B., Bailey, Andy M., Foster, Gary D., Mills, P. R. and Challen, Michael P. (2009) Quantifiable downregulation of endogenous genes in Agaricus bisporus mediated by expression of RNA hairpins. Journal of Microbiology and Biotechnology, Vol.19 (No.3). pp. 271-276. doi:10.4014/jmb.0806.398

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Official URL: http://dx.doi.org/10.4014/jmb.0806.398

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Abstract

Functional gene studies in the cultivated white button mushroom Agaricus bisporus have been constrained by the absence of effective gene-silencing tools. Using two endogenous genes from A. bisporus, we have tested the utility of dsRNA hairpin constructs to mediate downregulation of specific genes. Hairpin constructs for genes encoding orotidine 5'-monophosphate decarboxylase (URA3) and carboxin resistance (CBX) were introduced into A. bisporus using Agrobacterium-mediated transfection. Although predicted changes in phenotype were not observed in vitro, quantitative-PCR analyses indicated unambiguously that transcripts in several transformants were substantially reduced compared with the non-transformed controls. Interestingly, some hairpin transformants exhibited increased transcription of target genes. Our observations show that hairpin transgenic sequences can mediate downregulation of A. bisporus endogenous genes and that the technology has the potential to expedite functional genomics of the mushroom.

Item Type: Journal Article
Subjects: T Technology > TP Chemical technology
Q Science > QR Microbiology
Divisions: Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) > Warwick HRI (2004-2010)
Journal or Publication Title: Journal of Microbiology and Biotechnology
Publisher: Han'gug Mi'saengmul Saengmyeong Gong Haghoe
ISSN: 1017-7825
Official Date: March 2009
Dates:
DateEvent
March 2009Published
Volume: Vol.19
Number: No.3
Number of Pages: 6
Page Range: pp. 271-276
DOI: 10.4014/jmb.0806.398
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
Funder: Great Britain. Dept. for Environment, Food & Rural Affairs (DEFRA), Biotechnology and Biological Sciences Research Council (Great Britain) (BBSRC)

Data sourced from Thomson Reuters' Web of Knowledge

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