Ding Yi-Xin, Ou-Yang Xiang, Shang Chang-Hua, Ren Ang, Shi Liang, Li Yu-Xiang, Zhao Ming-Wen
College of Life Sciences, Nanjing Agricultural University, Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture, Jiangsu, PR China.
Biosci Biotechnol Biochem. 2008 Jun;72(6):1571-9. doi: 10.1271/bbb.80067. Epub 2008 Jun 7.
A farnesyl-diphosphate synthase gene, designated GlFPS, was isolated from a triterpene-producing basidiomycetous fungus, Ganoderma lucidum. The GlFPS cDNA was found to contain an open reading frame of 1,083 bp, encoding a protein of 360 amino acids with a calculated molecular mass of 41.27 kDa. The deduced amino acid sequence of the GlFPS cDNA exhibited a high homology with other fungal FPS genes, and contained four conserved domains. Phylogenetic analysis showed that GlFPS belonged to the basidiomycete FPS group. Competitive PCR revealed that GlFPS was constitutively expressed in the mycelium growth stage, whereas the transcripts of GlFPS accumulated to high levels rapidly during the process of mushroom primordia. Treatment of mycelia with exogenous methyl jasmonate also caused a large accumulation of GlFPS mRNA. Subsequently, promoter analysis indicated that the 5' upstream region of GlFPS possessed various potential regulatory elements associated with physiological and environmental factors. Functional complementation of GlFPS in an ERG20-disrupted yeast strain indicated that the cloned cDNA encoded a farnesyl-diphosphate synthase.
从产三萜的担子菌灵芝中分离出一个法尼基二磷酸合酶基因,命名为GlFPS。发现GlFPS cDNA包含一个1083 bp的开放阅读框,编码一个360个氨基酸的蛋白质,计算分子量为41.27 kDa。GlFPS cDNA推导的氨基酸序列与其他真菌FPS基因具有高度同源性,并包含四个保守结构域。系统发育分析表明,GlFPS属于担子菌FPS组。竞争性PCR显示,GlFPS在菌丝体生长阶段组成型表达,而在蘑菇原基形成过程中,GlFPS的转录本迅速积累到高水平。用外源茉莉酸甲酯处理菌丝体也会导致GlFPS mRNA大量积累。随后,启动子分析表明,GlFPS的5'上游区域具有与生理和环境因素相关的各种潜在调控元件。在ERG20基因缺失的酵母菌株中对GlFPS进行功能互补表明,克隆的cDNA编码一种法尼基二磷酸合酶。