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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (05): 55-60    DOI:
    
Suppression of Taxoid 14β-hydroxylase Gene Expression in Taxus × media via RNA Interference
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Abstract  Taxoid 14β-hydroxylase directs a side-route of taxol pathway that leading to a large number of 14β-hydroxy taxoid side products. A RNAi vector was used to suppress the expression of taxoid 14β-hydroxylase (14OH) gene in Taxus×media suspension cells. The dsRNA expression unit was introduced into Taxus×media cells via Agrobacterium tumefaciens-mediated transformation approach. PCR-Southern blot analysis of transgenic cells showed that the dsRNA unit was integrated into the genome of Taxus×media. The levels of transcripts for 14OH gene in transformed and the control cell lines were determined by using RT-PCR method. In addition, the yield of taxoids were analyzed by HPLC. The transcription of 14OH gene was suppressed and the total amount of three C-14 oxygenated taxoids (yunnanxane, sinenxan A, sinenxan C) in Taxus×media transgenic cell lines reduced significantly as compared to those in the non-transgenic control. Inhibition of the 14OH gene via RNAi might provide a new way to enhance the production of Taxol in Taxus cells.

Key wordsRNA interference;Gene expression;Taxus x media;Taxoid 14β-hydroxylase     
Received: 19 December 2008      Published: 06 May 2009
Cite this article:

LI Feng-Lan- Qiu-De-Wei- Ma-Xiao-Jun- Liu-Min- Hu-De-Rong. Suppression of Taxoid 14β-hydroxylase Gene Expression in Taxus × media via RNA Interference. China Biotechnology, 2009, 29(05): 55-60.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I05/55

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