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首页> 美国卫生研究院文献>Applied and Environmental Microbiology >Identification and Expression Analyses of Putative Sesquiterpene Synthase Genes in Phormidium sp. and Prevalence of geoA-Like Genes in a Drinking Water Reservoir
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Identification and Expression Analyses of Putative Sesquiterpene Synthase Genes in Phormidium sp. and Prevalence of geoA-Like Genes in a Drinking Water Reservoir

机译：拟南芥中倍半萜合酶基因的鉴定与表达分析饮用水水库中geoA-like基因的克隆与流行

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摘要

The occurrence of taste and odor problems in drinking water supplies is a widespread phenomenon. From a Saxonian water reservoir we isolated a cyanobacterial species which was classified as Phormidium sp. Under laboratory conditions it produced an earthy-musty smell due to the synthesis of geosmin. The only genes shown to be involved in geosmin biosynthesis are cyc2 and geoA of Streptomyces. Based on the alignment of Cyc2 with a putative sesquiterpene synthase of Nostoc punctiforme, a degenerate primer pair was designed. By PCR, we could amplify two similar genes in Phormidium sp., which we named geoA1 and geoA2. Their expression was studied by reverse transcription-PCR. This revealed that both genes are expressed at 20°C and a light-dark cycle of 12 h. Expression was not detectable at the end of a 24-h dark period. To analyze the prevalence of geoA1 and geoA2 in samples from the phytobenthos, we generated PCR fragments with the same degenerate primer pair. Fifty-five different sequences that might represent geoA variants were obtained. The GC content ranged from 42% to 67%, suggesting that taxonomically very different bacteria might contain such genes.
机译：饮用水供应中的味道和气味问题的发生是普遍现象。从萨克森州的水库中，我们分离出一种蓝细菌，其被分类为Phormidium sp。在实验室条件下，由于合成了土臭素，它产生了泥土味。唯一被证明参与土工生物合成的基因是链霉菌的cyc2和geoA。基于Cyc2与点状的Nostoc点状倍半萜合酶的比对，设计了简并引物对。通过PCR，我们可以在Phormidium sp。中扩增两个相似的基因，我们将其命名为geoA1和geoA2。通过逆转录PCR研究它们的表达。这表明这两个基因均在20°C和12h的明暗循环中表达。在24小时黑暗时段结束时无法检测到表达。为了分析来自植物底栖动物的样品中geoA1和geoA2的患病率，我们生成了具有相同简并引物对的PCR片段。获得了可能代表geoA变体的55种不同序列。 GC含量在42％至67％之间，表明在分类学上非常不同的细菌可能含有此类基因。

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